SUPPLEMENTARY INFORMATION
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- Daniel Merritt
- 5 years ago
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Transcription
1 o:./ntur mrna o/o vs. wt STD TTEST MIR-A 9,,7797,E- MIR--,99,7 MIR-7,7,7,9 MIR-B,99 79,9 MIR-E,97,77,99 MIR-7,9 99,99 LET-7D,77,77,7 MIR-,9979 7,99 MIR-D,99,7, MIR-9,7 9,99 LET-7F,7,9,7 MIR-9 7,9777,7 MIR-A 79999,979,99 MIR-7A 77 99, MIR-7B 7,, LET-7C 9,9,799 MIR- 7,79,99 MIR-A,7797,77,7 MIR-,,79,97 MIR-C,9,,99 LET-7G,9,9,97 MIR-9C,7,,9 MIR-9B,99,7,77 MIR-A,,79,9 LET-7B 977,97,99 MIR- 97,99,9 MIR-A,7, mrna o/o vs. wt STD TTEST MIR-A,9977, MIR-D-P,799 MIR-B-P 97,,99 MIR-E,B,O-P 799,7797,799 MIR-,97,9,7 MIR-,77,79,7 MIR--P,77,9,79 MIR-B-P,977,7,7 MIR-9B,79,79,9 MIR--MIR-,9797,,977 MIR-A,779,9,9997 MIR-7,77,977,979 MIR-,777,77,7 MIR-99 99,77,77 MIR-A-P 7,7,7 MIR- 7,9,9 MIR-9B-P 7,777, MIR-A-P 77,797,9 MIR-7,779,7 MIR-9-P 779,9777,99 MIR-B,99,9 MIR-7B-7C,7,9,97 MIR-7C,77,77,9E- MIR-,9,, MIR-B,7,79,E- MIR-99-P,7,7 7,9E- MIR-7-P,9,, MIR-7B,77,799,77E- MIR ,97797,9 MIR-7-P 77779,997,E- mrna DIO vs. how STD TTEST MIR-A,79 999,7 MIR-B 79,7,9 MIR-,7 977,9 MIR-7,,97,9 MIR--,79,7 MIR-7,7,7 LET-7D,9997,9,7 LET-7F,97,9,7 MIR-9B,,7,7 MIR--P 9,7, LET-7C 9,79,9 MIR-7A 799,79,97 LET-7A 97, LET-7G 77,779,97979 MIR-B 97 7, MIR-E 7,79, MIR-A,999,9977,7 MIR-9,9,7799,7 LET-7B,7,99,7 MIR-D,7,7,797 MIR-9C,7979,779,7 MIR-9,7,97,79 MIR-,97,9, MIR-B,7,9, MIR-C,,97,97 MIR-,,99, MIR-,,7,77 MIR-7B 77,7777,997 MIR-B 7,7,7 MIR-A 97,7,9 MIR-A 77,99,79 MIR- 7,77,9 MIR-A 9,79, Supplmntry Tl (, ) Lst of > fol up-rgult mrnas (p<, twn thnl quruplts, n p<, of th ologl trplts wth vrg sgnl ntnsty of fol ovr kgroun) n os o/o (), n DIO () vs. g mth C7Bl/ ontrol m. () Lst of < fol own-rgult mrnas (p<, twn thnl quruplts, n p<, of th ologl trplts). Th shown folrgulton, stnr vton (STD) n p- vlus wr lult from th ologl trplts.
2 BW Pl. Gl. GINF R lmp R. Stmul. B.Gl.Clr. (g) (mg/l) (mg/kg.m) (mg/kg.m) (Fol) (ml/kg.mn) PBS,±,7 ± 9,±,,9±, ±,,± Ant- 9,±, 9,7±9,,±, ±,7,± ±, lmp HGP HGP sup. DOG Gstro DOG Hrt DOG BAT DOG WAT (mg/kg.m) (%) R(ng/mg/mn) R(ng/mg/mn) R(ng/mg/mn) R(ng/mg/mn) PBS,± ±9,,±, 7,±, ±, ±, Ant- 7,±,,±,±, ±,,±, ± Supplmntry Tl Hyprnsulnm-uglym lmp stus n nt-, or PBS-njt o/o m (n=7).
3 Syn. mr Syn. mr MR- nm nm n o s s n o s s r o n s s. x p r 7 - l r. x p - l p. v x. r m R ) s l l mr-: ' A G TATCGGGACATGTTACGACGA ' mr-7: ' A C TATCGGGACATGTTACGACGA ' u o n ( Syn.mR-7 (. nm) Syn.mR- (. nm) R l T m 7 R l T m R. M R. M R, wt o/o Ctrl DIO R, wt o/o Ctrl DIO Supplmntry Fgur () Northrn lottng for mr- on,, or nm syntht mr- or mr-7. () Rltv mr-, or mr-7 lvls msur y quntttv rl-tm PCR usng, nm syntht mr-, or mr-7 tmplts s nt. (, ) Rltv mr- (), or mr-7 () xprsson lvls n lvrs of wltyp (wt), o/o, how or DIO m s nt, normlz to U RNA lvls. Mns ± s.. r shown for ll pnls.
4 Rl. mrna xprsson lvls, Control HBV HCV Ctrl+HBP+HCV ASH NAFLD NASH Sujts (n) mr- mr-7 Rltv mr-7 xprsson,., HOMA R =, p, Rl. mrna xprsson lvls HOMA <, HOMA >, mr- mr-7 n=9 ±STE Supplmntry Fgur () Rltv mr-, or mr-7 lvls n lvr opss of hlthy (n=), hron hptts B n C vrus (HBV, n=7, n HCV n=7)-nft nvuls n humn ptnts wth lohol sttohptts (ASH, n=7), non-lohol ftty lvr ss (NAFLD, n=) n wth non-lohol sttohptts (NASH, n=) quntf y rl tm PCR, normlz to U. () Corrlton (n=9) twn th rltv mr-7 lvls s quntf y rl tm PCR, normlz to U, n th HOMA-IR nx of hlthy (n=), hron hptts B n C vrus (HBV, n=, n HCV n=7)- nft nvuls, n humn ptnts wth lohol sttohptts (ASH, n=), non-lohol ftty lvr ss (NAFLD, n=) n wth non-lohol sttohptts (NASH, n=). () Rltv mr- (lft), or mr-7 (rght) lvls n lvr opss of ptnts (n=9) s n () wth HOMA-IR nx low, or ovr. Brs nt mn ± SEM.
5 mr-7 A-GFP A-7/GFP mr-7 Dy Dy 7 Dy Dy 7 mr- mr- mr- Totl RNA Totl RNA -mr7/gfp PBS PBS Ant-7 Ant- PBS Ant. Ant.MM Ant. Ant.MM Ant. mr- mr- mr-7 mr- o/o Totl RNA Dy Dy 7 Dy Supplmntry Fgur () Northrn lottng for mr-, mr-, n mr- of µg of totl RNA from lvrs of C7Bl/J m njt wth -GFP, or -7/GFP. () Northrn lottng for mr-7 usng µg of totl RNA from lvrs of C7Bl/J m njt wth -7/GFP or PBS. () Northrn lottng for mr-, mr-7, n mr- from lvrs of o/o m njt wth PBS, nt-, or nt-7. () Northrn lottng for mr-, n mr-7 usng µg of totl RNA from ft of o/o m njt wth PBS, nt-mm, or nt- t nt ys postnjton. Ethum rom (EtBr) stnng n (,, n ) s shown s long ontrol.
6 Ant. MM 7 Ant. o/o 7 Ant. MM 7 Ant. o/o Volum (ml/kg/hr) Tmprtur ( C) V(O ) V(O ) Rst V(CO ) V(CO ) Rst Dy (rnom) Dy (h fst) Foo (g) 7 Ant. MM 7 Ant. o/o Rltv gn xprsson,, Foo Foo / mss Cptα Aox MCAD VLCAD A A Supplmntry Fgur (-) Mtol g t of o/o m (n=) njt wth nt-mm, or nt- t y post njton. Rst: msurmnts urng rstng pros. () Rltv gn xprsson msur y rl-tm PCR n totl RNA from ft of o/o m s n (-) normlz to Gph. Mns ± s.. r shown for ll pnls.
7 Lp-nt-MM Lp-nt- MR- trna Lvr 9 Lp-nt-MM Lp-nt- MR- trna MR- Musl Ft Bloo gluos(mm) 7 trna Dy - - fst IPGTT Gluos(mM) Lp-nt-MM Lp-nt- ITT Gluos(mM) Lp-nt-MM Lp-nt- PTT Gluos(mM) Tm (mn) Lp-nt-MM Lp-nt- Tm (mn) f Insuln (ng/ml) Lp-nt-MM Lp-nt- DIO ( wks) h fst h rf Tm (mn) Supplmntry Fgur () Northrn lottng for mr- on µg of totl RNA from lvr, musl, or ft of DIO m njt wth µg/kg Lp-nt-, or Lp-nt-MM. Ethum rom (EtBr) stnng s shown s ontrol. () Bloo gluos lvls n rnom or fstng ontons n DIO m njt wth Lp-nt-MM, or Lp-nt- (n=). (-) I. p. gluos tolrn tst (IPGTT)(), U/kg.p. nsuln tolrn tst (ITT)(), n pyruvt tolrn tst of DIO m s n (). In (), vlus n th mn tm pont r normlz to. (f) Insuln lvls of DIO m s n () ftr h fst (lft) follow y h r-fng (rght). Mns ± s.. r shown for ll pnls. 7
8 BW (g) SC poyt sz struton (normlz to ll numr) Ant-MM7 Ant ,,,,,,,, CT Dy Dy Dy Dy Dy Dy Ant-MM7 Ant- o/o Orgn wght (g). Ant-MM, Ant-.,., Ingunl ft ps V poyt sz struton (normlz to ll numr),,,,,,, Lvr Qups Hrt Kny Spln Ant-MM7 Ant- o/o Apoyt sz Apoyt sz, f, SC poyt sz struton (normlz to ll numr),,,,,,,, Ant-MM7 Ant- DIO V poyt sz struton (normlz to ll numr),,,,,,, Ant-MM7 Ant- DIO Apoyt sz Apoyt sz Supplmntry Fgur () Boy wght (BW) of o/o m njt wth nt-, or nt-mm (n=). () Orgn wghts (g) of o/o m s n (). Mns ± s.. r shown for ll pnls. (-f) Cll sz proflng of poyts from suutnous (SC) (, ), n vsrl (V) (, f) ft of o/o (, ), n DIO m (, f) njt wth nt-, or nt-mm. Th numr of lls n h frton (/ n o/o, or / n DIO) ws v y th totl numr of nlyz lls. Brs show mn of th orrsponng frton from h nml ±STD (o/o n=, DIO n=).
9 Rl. mr7 xprsson, A-GFP A-mR-7 Bloo gluos (mm) 9 7 A-GFP A-mR-7 Insuln (ng/ml) A-GFP A-mR-7 VISC.FAT LIVER Dy (rnom) Dy (h fst) Dy 9 (rnom) Dy 9 (h fst) h fst f IPGTT (mm) Tm (mn) A-GFP A-7/GFP ITT Bloo gluos (%) A-GFP A-7/GFP Tm (mn) A-GFP HOMA-IR 7 A-7/GFP g Avr. poyt sz 9 7 A-GFP A-7/GFP 9 7 Ingunl SC Supplmntry Fgur 7 () Rltv mr-7 lvls n V ft (lft), or lvrs (rght) of C7Bl/J m njt wth -GFP, or -7/GFP followng surglly njton n th ngunl ft ps (n=7+), n normlz to U. (-f) Rnom n fstng loo gluos () n nsuln () lvls, IPGTT (), n ITT (), n HOMA-IR nx (f) of C7Bl/J m s n (). (g) Automt CllProflr quntfton of th vrg poy sz. 9
10 GGGAGAAACGTTCTCACTCGCTCTCTGCTCGCTGCGGGCGCTCCCCGCCCTCTGCTGCCAGAACCTTGGGGATGTGCCTAGACCCGGCGCAGCACACGTCCGGGCCAACCGCGAGCAGAACAAACCTTTGGCGGGCGGCCAGGAGGCTCCCTCCCAGCCACCGCCCCCCTCCAGCGCCTTTTTTTCCCCCCATACAATACAAGATCTTCCTTCCTCAGTTCCCTTAAAGCACAGCCCAGGGAAACCTCCTCACAGTTTTCATCCAGCCACGGGCCAGCATGTCTGGGGGCAAATACGTAGACTCGGAGGGACATCTCTACACCGTTCCCATCCGGGAACAGGGCAACATCTACAAGCCCAACAACAAGGCCATGGCAGACGAGCTGAGCGAGAAGCAAGTGTACGACGCGCACACCAAGGAGATCGACCTGGTCAACCGCGACCCTAAACACCTCAACGATGACGTGGTCAAGATTGACTTTGAAGATGTGATTGCAGAACCAGAAGGGACACACAGTTTTGACGGCATTTGGAAGGCCAGCTTCACCACCTTCACTGTGACGAAATACTGGTTTTACCGCTTGCTGTCTGCCCTCTTTGGCATCCCGATGGCACTCATCTGGGGCATTTACTTCGCCATTCTCTCTTTCCTGCACATCTGGGCAGTTGTACCATGCATTAAGAGCTTCCTGATTGAGATTCAGTGCATCAGCCGTGTCTATTCCATCTACGTCCACACCGTCTGTGACCCACTCTTTGAAGCTGTTGGGAAAATATTCAGCAATGTCCGCATCAACTTGCAGAAAGAAATATAAATGACATTTCAAGGATAGAAGTATACCTGATTTTTTTTCCTTTTAATTTTCCTGGTGCCAATTTCAAGTTCCAAGTTGCTAATACAGCAACAATTTATGAATTGAATTATCTTGGTTGAAAATAAAAAGATCACTTTCTCAGTTTTCATAAGTATTATGTCTCTTCTGAGCTATTTCATCTATTTTTGGCAGTCTGAATTTTTAAAACCCATTTAAATTTTTTTCCTTACCTTTTTATTTGCATGTGGATCAACCATCGCTTTATTGGCTGAGATATGAACATATTGTTGAAAGGTAATTTGAGAGAAATATGAAGAACTGAGGAGGAAAAAAAAAAAAAAGAAAAGAACCAACAACCTCAACTGCCTACTCCAAAATGTTGGTCATTTTATGTTAAGGGAAGAATTCCAGGGTATGGCCATGGAGTGTACAAGTATGTGGGCAGATTTTCAGCAAACTCTTTTCCCACTGTTTAAGGAGTTAGTGGATTACTGCCATTCACTTCATAATCCAGTAGGATCCAGTGATCCTTACAAGTTAGAAAACATAATCTTCTGCCTTCTCATGATCCAACTAATGCCTTACTCTTCTTGAAATTTTAACCTATGATATTTTCTGTGCCTGAATATTTGTTATGTAGATAACAAGACCTCAGTGCCTTCCTGTTTTTCACATTTTCCTTTTCAAATAGGGTCTAACTCAGCAACTCGCTTTAGGTCAGCAGCCTCCCTGAAGACCAAAATTAGAATATCCATGACCTAGTTTTCCATGCGTGTTTCTGACTCTGAGCTACAGAGTCTGGTGAAGCTCACTTCTGGGCTTCATCTGGCAACATCTTTATCCGTAGTGGGTATGGTTGACACTAGCCCAATGAAATGAATTAAAGTGGACCAATAGGGCTGAGCTCTCTGTGGGCTGGCAGTCCTGGAAGCCAGCTTTCCCTGCCTCTCATCAACTGAATGAGGTCAGCATGTCTATTCAGCTTCGTTTATTTTCAAGAATAATCACGCTTTCCTGAATCCAAACTAATCCATCACCGGGGTGGTTTAGTGGCTCAACATTGTGTTCCCATTTCAGCTGATCAGTGGGCCTCCAAGGAGGGGCTGTAAAATGGAGGCCATTGTGTGAGCCTATCAGAGT -AGCAGCATTGTACAGGGCTATGA- ATGTCTGGGGGCAAATACGTAGACTCGGAGGGACATCTCTACACCGTTCCCATCCGGGAACAGGGCAACATCTACAAGCCCAACAACAAGGCCATGGCAGACGAGCTGAGCGAGAAGCAAGTGTACGACGCGCACACCAAGGAGATCGACCTGGTCAACCGCGACCCTAAACACCTCAACGATGACGTGGTCAAGATTGACTTTGAAGATGTGATTGCAGAACCAGAAGGGACACACAGTTTTGACGGCATTTGGAAGGCCAGCTTCACCACCTTCACTGTGACGAAATACTGGTTTTACCGCTTGCTGTCTGCCCTCTTTGGCATCCCGATGGCACTCATCTGGGGCATTTACTTCGCCATTCTCTCTTTCCTGCACATCTGGGCAGTTGTACCATGCATTAAGAGCTTCCTGATTGAGATTCAGTGCATCAGCCGTGTCTATTCCATCTACGTCCACACCGTCTGTGACCCACTCTTTGAAGCTGTTGGGAAAATATTCAGCAATGTCCGCATCAACTTGCAGAAAGAAATATAA ACAAGATCTTCCTTCCTCAGTTCTCTTAAATCACAGCCCAGGGAAACCTCCTCAGAGCCTGCAGCCAGCCACGCGCCAGCATGTCTGGGGGCAAATACGTAGACTCCGAGGGACATCTCTACACTGTTCCCATCCGGGAACAGGGCAACATCTACAAGCCCAACAACAAGGCCATGGCAGACGAGGTGACTGAGAAGCAAGTGTATGACGCGCACACCAAGGAGATTGACCTGGTCAACCGCGACCCCAAGCATCTCAACGACGACGTGGTCAAGATTGACTTTGAAGATGTGATTGCAGAACCAGAAGGGACACACAGTTTCGACGGCATCTGGAAGGCCAGCTTCACCACCTTCACTGTGACAAAATATTGGTTTTACCGCTTGTTGTCTACGATCTTCGGCATCCCAATGGCACTCATCTGGGGCATTTACTTTGCCATTCTCTCCTTCCTGCACATCTGGGCGGTTGTACCGTGCATCAAGAGCTTCCTGATTGAGATTCAGTGCATCAGCCGCGTCTACTCCATCTACGTCCATACCTTCTGCGATCCACTCTTTGAAGCTATTGGCAAGATATTCAGCAACATCCGCATCAGCACGCAGAAAGAGATATGAGGGACATTTCAAGGATGAAAGGTTTTTTTCCCCCCTTACTATTTCCTTGGTGCCAATTCCAAGTTGCTCTCGCAGCAGCAAATTTATGAATGGTTTGTCTTGATCAAGAACAAAGAATTCATTCCCACCATTCTCATATATACTACTTGTCTCTTCTAAGCTACTGCATCTATGTTTGACAGTCTGGAATGTTTAAACCCATTCCTGCTCTCTCTTTTATATGTGAATCATTGTTTCATTGGCTAAAATATAAACATATTGTTGAAAGATGATTTGAGAAAAATAGGAAGGACTGGGAGGCAGGGAAGAGTACCAACAACCTCAACTGCCTACTCAAAGGTGATGATGTCATACAAAGGGAAGAGATTCAGGTTACGGCCATTTGTTTAGGGGCATGAAGGAACGTTTTTAATATATGCCAGTTATCTAAGGAATTGGTTGCTGTCCTCACTCTTAACAATCCAGTTAGATTTAGGGATTTAGGGATCACCATCAATTTGGAGACTATAATCTTCATGATACCAACAATGTTTTACTTATCCTGGCATTTTAACCTGTTATTTTGTATGCCTGAATATTTGCTATACTGAGAATAAGACCTACGTGCCTTCTAATTTTTCATGTTTTTTTTTTTTCCAAATAGGATCTAACTCATCTACTTGCATGATGCCGGCAGCTTTCCTAAAAACAAAACATACAAATTGCACTTGCTAGTTCTCTGTACTTGTTTCTGACTCTGAAATACAGAACCTGTTGATGTTGATATCTGTGCTCAGCTATGTAGCATCTTTCTCTCTGTTAAGCCTGGTCAACATTAACCCAATGAAATGATTTGAAGCAGACAAATGGGGGTGAGACCTCTCTGGACTGGCAGAAGTAGAAGCCAGCTTTCCCTGCCACTCAGCAACTGAATGAGGCCAGCGTGTCTATTCAGTTTCACTCATTTTCAAGAATAATCACAGGTTCCTGACTCTAAGCCAGCCCCTCACCAGGATCAAGGTTTAGTGACTGACTGGGATGATTTAGGAGCTCAACATTGTACTTCCTTTTCAGCTGATGAGTGAACCTCCAGGGAGGGGTGTCAAAAAGGAG ATGTCTGGGGGCAAATACGTAGACTCCGAGGGACATCTCTACACTGTTCCCATCCGGGAACAGGGCAACATCTACAAGCCCAACAACAAGGCCATGGCAGACGAGGTGACTGAGAAGCAAGTGTATGACGCGCACACCAAGGAGATTGACCTGGTCAACCGCGACCCCAAGCATCTCAACGACGACGTGGTCAAGATTGACTTTGAAGATGTGATTGCAGAACCAGAAGGGACACACAGTTTCGACGGCATCTGGAAGGCCAGCTTCACCACCTTCACTGTGACAAAATATTGGTTTTACCGCTTGTTGTCTACGATCTTCGGCATCCCAATGGCACTCATCTGGGGCATTTACTTTGCCATTCTCTCCTTCCTGCACATCTGGGCGGTTGTACCGTGCATCAAGAGCTTCCTGATTGAGATTCAGTGCATCAGCCGCGTCTACTCCATCTACGTCCATACCTTCTGCGATCCACTCTTTGAAGCTATTGGCAAGATATTCAGCAACATCCGCATCAGCACGCAGAAAGAGATATGA TGCTGCAAACCTGACCCCTGCTCAGTAAAGCACTTGCAACCGTCTGTTATGCTGTGACACATGGCCCCTCCCCCTGCCAGGAGCTTTGGACCTAATCCAAGCATCCCTTTGCCCAGAAAGAAGATGGGGGAGGAGGCAGTAATAAAAAGATTGAAGTATTTTGCTGGAATAAGTTCAAATTCTTCTGAACTCAAACTGAGGAATTTCACCTGTAAACCTGAGTCGTACAGAAAGCTGCCTGGTATATCCAAAAGCTTTTTATTCCTCCTGCTCATATTGTGATTCTGCCTTTGGGGACTTTTCTTAAACCTTCAGTTATGATTTTTTTTTCATACACTTATTGGAACTCTGCTTGATTTTTGCCTCTTCCAGTCTTCCTGACACTTTAATTACCAACCTGTTACCTACTTTGACTTTTTGCATTTAAAACAGACACTGGCATGGATATAGTTTTACTTTTAAACTGTGTACATAACTGAAAATGTGCTATACTGCATACTTTTTAAATGTAAAGATATTTTTATCTTTATATGAAGAAAATCACTTAGGAAATGGCTTTGTGATTCAATCTGTAAACTGTGTATTCCAAGACATGTCTGTTCTACATAGATGCTTAGTCCCTCATGCAAATCAATTACTGGTCCAAAAGATTGCTGAAATTTTATATGCTTACTGATATATTTTACAATTTTTTATCATGCATGTCCTGTAAAGGTTACAAGCCTGCACAATAAAAATGTTTAACGGTT GCTGCTAAACCGAGACTGCCAAGCCTGTTGTAAACATGACCCCTTTTATGCAAAGCCCTTGCAATAGTCTGCATGCTGTGAAGCTCGACCTTTCCCCCTGCAAGGAACCTTTGACCTAATCCAACCATCATTTTGTTCAGAAAGGTGGGGGAAGGGTGGTAACAAAAGCTTGAGGTAATGTTCTTGCTGTAATAAATTCAAGTTTTTCTGAACCCAAACTGAGGAATTTCACCTGTGTACCTGAGTCTCCAGAAAGCTGCCTGCCTGGGACACCCAAAAGCCTTTTACTTCCCAGCTCACATTACAGCTCTGCCCTTGGGGATATTTTTAAAATTCCAGATAGGCTTTCATTTTCACTTTCATACATGTATTGGAACCCTGCTTGACTTGTTTTCTCCTTCAGTCTTGCCGACACTTTACCAACCTGCTACCTACTTTGATTGTTTGCATTTAAAACAGACACTGGCATGGCCACAGTTTGAATTTTAAACTGTGCACATAACTGAAAGTGTACTAGACTGTATACCTTTTTACATGTAGAGATATTCTTTATCTTTATATAAGGAGAATCACTTGGGAAATGATTCTACAATTCAGTCTGTAAACTGTGTGTTCCAAGACATGTCTGTTCTCCCTAGATACTCAGTTTTATACAAGTCAATTGCTGATCCAAAAGGTTACTGAAATTTTATATGCTTACTGATATATTTTACACTTTTTT ATGCTGCATGTCCTATAAAGATTTCAAATCTGCACAATAAAATTGTTTAACAGTTA -AGCAGCATTGTACAGGGCTATGA- AGCCTGGTCAACATTAACCCAATGAAATGATTTGAAGCAGACAAATGGGGGTGAGACCTCTCTGGACTGGCAGAAGTAGAAGCCAGCTTTCCCTGCCACTCAGCAACTGAATGAGGCCAGCGTGTCTATTCAGTTTCACTCATTTTCAAGAATAATCACAGGTTCCTGACTCTAAGCCAGCCCCTCACCAGGATCAAGGTTTAGTGACTGACTGGGATGATTTAGGAGCTCAACATTGTACTTCCTTTTCAGCTGATGAGTGAACCTCCAGGGAGGGGTGTCAAAAAGGAGGCTGCTAAACCGAGACTGCCAAGCCTGTTGTAAACATGACCCCTTTTATGCAAAGCCCTTGCAATAGTCTGCAATGCTGTGAAGCTCGACCTTTCCCCCTGCAAGGAACCTTTGACCTAATCCAACCATCATTTTGTTCAGAAAGGTGGGGGAAGGGTGGTAACAAAAGCTTGAGGTAATGTTCTTGCTGTAATAAATTCAAGTTTTTCTGAACCCAAACTGAGGAATTTCACCTGTGTACCTGAGTCTCCAGAAAGCTGCCTGCCTGGGACACCCAAAAGCCTTTTACTTCCCAGCTCACATTACAGCTCTGCCCTTGGGGATATTTTTAAAATTCCAGATAGGCTTTCATTTTCACTTTCATACATGTATTGGAACCCTGCTTGACTTGTTTTCTCCTTCAGTCTTGCCGACACTTTACCAACCTGCTACCTACTTTGATTGTTTGCATTTAAAACAGACACTGGCATGGCCACAGTTTGAATTTTAAACTGTGCACATAACTGAAAGTGTACTAGACTGTATACCTTTTTACATGTAGAGATATTCTTTATCTTTATATAAGGAGAATCACTTGGGAAATGATTCTACAATTCAGTCTGTAAACTGTGTGTTCCAAGACATGTCTGTTCTCCCTAGATACTCAGTTTTATACAAGTCAATTGCTGATCCAAAAGGTTACTGAAATTTTATATGCTTACTGATATATTTTACACTTTTTTATGCTGCATGTCCTATAAAGATTTCAAATCTGCAC AGCAGCATTGTACAGGGCTATGA hcv s (Lvr) xprsson Rl.,., TGCTGCA hs hs GCTGCT ATGCTG PBS Ant- o/o Cv Gpnm Prom Lpl Pl Lypl ApoB BCKDHA S S Ln ATGCTGCA TGCTGCA ATGCTG Humn mcv mous rt opossum og ow humn hmp rhsus hkn s TATCGG ACA ms ms ms GTTACGACG Mous GCTGCT CAATGCTG ATGCTGCA Supplmntry Fgur Gn xprsson nlyss mr-/7 trgt gns. () Mrorry nlyss: Assssmnt of hngs n gn xprsson rtly nu y ovr-xprsson (A-7) of mr-/7. Gns rryng s mth to mr-/7 n th ' UTR r rprsnt y th grn urv, th sust of gns hrorng s lkly to unr volutonry sltv prssur s shown n r, whl th lk ln ounts for gns wth no s mth ovr th ntr mrna squn. Dots nt th mn log-fol hng of th thr sts of gns. Th p-vlus rport on th thr pnls ompr th grn n r urvs to th lk urv wth on-s Wloxon rnk-sum tst. () Gn Ontology trms ovrrprsnt n th top gns puttvly trgt y mr-/7. () Hpt gn xprsson nlyss y rl tm PCR of o/o m njt wth PBS or nt- (n=). () Grphl rprsntton of Cv- ong squn (CDS) n t s UTR, nlung th s motfs of humn n mous trnsrpts. Th s squns r mrk n r, n th mthng rsus twn Cv- UTR n th mr- proxml to th s squn n r. () Multpl lgnmnts of Cv- s UTR rgon n n slo hyrzton of mr-7 to th onsrv Cv- mrna nng st. Mns ± s.. r shown.
11 Bloo gluos (mm) Cv-KO A-GFP Cv-KO A-7/GFP C7Bl, A-GFP C7Bl, A-7/GFP Tm (mn) Bloo gluos (normlz vlus) Cv-KO A-GFP Cv-KO A-7/GFP C7Bl, A-GFP C7Bl, A-7/GFP Tm (mn) Bloo gluos (mm) Cv-KO A-GFP Cv-KO A-7/GFP C7Bl, A-GFP C7Bl, A-7/GFP Tm (mn) 9 A-GFP A-7/GFP Cv-KO Ant-MM Ant- Bloo gluos (mm) 7 Bloo gluos (mm) 7 9 o/n fst o/n fst Dy 7 9 f g h IPGTT Gluos (mm) Ant-MM Ant- ITT Gluos (norm. vlus) Ant-MM Ant- PTT Gluos (mm) Ant-MM Ant- Tm (mn) Tm (mn) Tm (mn) Rltv gn xprsson, Ant-MM Ant-, Cptα MCAD VLCAD Aox Aox Supplmntry Fgur 9 (-) Effts of gn-of- mr-7 funton (-) n mr- slnng (-) n Cv-KO m. IPGTT (),,7 U/kg ITT (), PTT () n -GFP, or -7/GFP tl vn njt C7Bl/J (lu n r, n=), or Cv-KO (grn n lk, n=) m. In (), vlus t th mn tm pont r normlz to. () Bloo gluos lvls n rnom or fstng ontons of Cv-KO m njt wth -GFP, or -7/GFP (n=). () Bloo gluos lvls n rnom or fstng ontons n Cv-KO m kpt on HFD for ys, njt wth Ant-MM, or Ant- (n=7). (f-h) IPGTT (f), U/kg ITT (g), n PTT (h), of DIO Cv-KO m s n (). In (g), vlus n th mn tm pont r normlz to. () Rltv gn xprsson msur y rl-tm PCR n totl RNA from ft of CvKO m f HFD for wks, n njt wth Ant-MM-, or Ant-. Exprsson ws normlz to Gph. Mns ± s.. r shown for ll pnls.
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