ampk ca mice  (Jackson Laboratory)

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Western blot analysis of female mouse livers fed HFD containing doxycycline (Doxy) to express constitutive active AMPK [a1(1-312); AMPK CA ] for 7 days compared to control livers. ( B ) Immunohistochemical analysis of livers from male control and AMPK CA mice fed HFD+Doxy for 7 months. Scale bars, 100 μm. Right: Quantification of the staining intensity for P-Thr 172 AMPK and P-Ser 79 ACC staining. n = 6 to 10. ±SEM, Welch t test. ( C ) Representative whole-mount (top) and MRI (bottom) image of male control and AMPK CA livers 9 months post–DEN injection. Yellow dashed lines indicate tumors. ( D ) Quantification of the tumor number in male control and AMPK CA livers 6 and 9 months post–DEN injection based on MRI images. 6 months: N = 5 to 7; 9 months: N = 16 to 17; Fisher least significant difference (LSD) test. ( E ) Quantification of the serum AFP levels in control and AMPK CA male mice 6 and 9 months post–DEN injection. n = 9 to 15. Fisher LSD test. * P < 0.05; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Western Blot, Control, Immunohistochemical staining, Staining, Injection

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Quantification of the tumor size from MRI imaging at 6 and 9 months. n = 10 to 448. Log normal Welch t test. ( B ) Immunohistochemical analysis of BrdU incorporation in tumor tissue from control and AMPK CA livers following a 4-hour BrdU pulse (10 mg/kg). Scale bars, 100 μm. Right: Quantification of the number of positive cells per area. n = 13 to 17. Welch t test. ( C ) Gene expression of Ampka1 normalized to Actin in nontumor and tumor tissue. n = 5 to 6. ±SEM, Fisher LSD test. ( D ) Immunohistochemical analysis of P-Ser 79 ACC in nontumor and tumor tissue from control and AMPK CA livers. Scale bars, 100 μm. ( E ) Quantification of the staining intensity of P-Ser 79 ACC. n = 5 to 14. Fisher LSD test. ( F ) Immunohistochemical analysis of P-Thr 172 AMPK in nontumor and tumor tissue from control and AMPK CA livers. Scale bars, 100 μm. ( G ) Quantification of the staining intensity. n = 6 to 11. Fisher LSD test. ( H ) Western blot analysis of nontumor (NT) and tumor (T) tissue. VINCULIN as loading control. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Imaging, Immunohistochemical staining, BrdU Incorporation Assay, Control, Gene Expression, Staining, Western Blot

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Weight of male control and AMPK CA mice on HFD for 8 and 16 weeks following STZ injection n = 14 to 19. Fisher LSD test. ( B ) Fasting blood glucose levels 20 weeks post–STZ injection. n = 6 to 8. Welch t test. ( C ) Oil Red O staining of control and AMPK CA livers 20 weeks following STZ injection. Right: Scoring of Oil Red O levels. n = 7. Welch t test. Scale bar, 100 μm. ( D ) Quantification of the triglyceride levels in livers of control and AMPK CA mice 20 weeks post–STZ injection. n = 5. Welch t test. ( E ) Whole-mount image (top) and MRI image (bottom) of livers 20 weeks following STZ injection. Yellow dashed lines indicate tumors. ( F ) Quantification of the tumor number from control and AMPK CA mice injected with STZ. n = 8. Welch t test. ( G ) Liver weight relative to body weight of STZ-injected mice. n = 13 to 14. Welch t test. ( H ) Quantification of tumor sizes 20 weeks following STZ injection. n = 67 to 136 from 13 to 14 mice. Log normal Welch t test. ( I ) Gene expression analysis of nontumor and tumor tissue from STZ-injected mice. n = 6 to 7. Fisher LSD test. ( J ) Western blot analysis of matched nontumor and tumor tissue from AMPK CA mice injected with STZ. Right: Quantification of total and phospho-AMPK CA levels. n = 10 to 11. Welsh t test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Control, Injection, Staining, Gene Expression, Western Blot

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Quantification of the relative levels of bile acids in livers from control and AMPK CA mice following 2 months (2 m) of doxycycline. n = 5. Welsh t test. ( B ) Heatmap of average relative bile acids levels of AMPK CA livers relative to control livers following varying durations of doxycycline treatment [15 days (15d) and 2 months] or in matched nontumor and tumor tissues [7 months (7 m) and “Tumor”], n = 4 to 6 per genotype per time point. ( C ) Ratio of conjugated versus nonconjugated bile acids in livers from control or AMPK CA mice on chow or HFD for 15 days. n = 4 to 6. Welsh t test. ( D ) Ratio of secondary (DCA;LCA) versus primary (CA;CDCA) bile acids in livers from control or AMPK CA mice on chow or HFD for 15 days. n = 4 to 6. Fisher LSD test. DCA, deoxycholic acid; LCA, lithocholic acid; CA, cholic acid; CDCA, chenodeoxycholic acid. ( E ) Quantification of the serum bile acid levels relative to control livers of tumor bearing mice on doxycycline diet for 7 months. n = 5 to 6. Welch t test. ( F ) Quantification of the TBARS concentration in livers from control or AMPK CA mice on chow or HFD containing doxycycline for 15 days or 2 months. n = 4 to 6. Fisher LSD test. ( G ) Quantification of the gene expression in control or AMPK CA livers of mice on chow or HFD containing doxycycline for 3 or 15 days or in tumors. n = 4 to 6. Fisher LSD test. ( H ) Western blot analysis of livers from mice on HFD containing doxycycline for 15 days. ( I ) Gene expression of YAP target genes in WT primary hepatocytes treated with 80 μM TLCA or DMSO for 6 hours. n = 5 to 6. Two-way analysis of variance (ANOVA). ( J ) Western blot analysis of primary hepatocytes treated with 80 μM TLCA or DMSO for 30 min, 1 hour, or 2 hours. VINCULIN as loading control. n.s., not significant.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Control, Concentration Assay, Gene Expression, Western Blot

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Gene expression of bile acid biosynthesis proteins and bile acid response genes in livers from mice fed HFD containing doxycycline for 15 days. n = 4. Fisher LSD test. ( B ) Heatmap of the log 2 fold change of gene expression changes in AMPK CA relative to control livers from mice on HFD containing doxycycline for 2 months. ( C ) Gene expression of bile acid transporters in livers from mice fed chow or HFD containing doxycycline for 15 days. n = 4 to 6. Fisher LSD test. ( D ) Gene expression of bile acid transcription factor in livers from mice fed chow or HFD containing doxycycline for 15 days. n = 4 to 6. Fisher LSD test. ( E ) Gene expression of bile acid responsive genes in the ileum from mice fed HFD containing doxycycline for 15 days. n = 4. Fisher LSD test. ±SEM. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Gene Expression, Control

Journal: Science Advances

Article Title: Constitutive AMPK activation prevents hepatocellular carcinoma development through inhibition of HNF4α activity

doi: 10.1126/sciadv.aea8017

Figure Lengend Snippet: ( A ) Transcription factors (TFs) enriched in AMPK CA mice relative to control mice following 2 months of HFD containing doxycycline using Enrichr and a 1.3-fold change with an adjusted P value less than 0.05. ( B ) Gene set enrichment analysis of genes in AMPK CA livers compared to control livers following 2 months of HFD containing doxycycline. NES, normalized enrichment score. ( C ) Western blot analysis of livers from control and AMPK CA mice following 15 days of HFD containing doxycycline. ( D ) Volcano plot of HNF4α binding sites altered in livers from control and AMPK CA mice following 15 days of HFD containing doxycycline. ( E ) Genomic location and directionality of change in HNF4α binding sites in livers from control and AMPK CA mice following 15 days of HFD containing doxycycline. TTS, transcriptional termination site. ( F ) Representative HNF4α binding sites in control versus AMPK CA livers. ( G ) Heatmap analysis of HNF4α target genes significantly changed (FDR < 0.05) in AMPK CA livers compared to control livers following 2 months of HFD containing doxycycline. Log 2 fold change. ( H ) Growth curve of the xenograft of MHCC-97H cells expressing WT or S304A or S304D mutant HNF4α. Inset: Representative image of tumor sizes at endpoint n = 11. ( I ) Weight of xenografts of MHCC-97H cells expressing WT or mutant HNF4α. n = 11. Welch t test. ( J ) Gene expression analysis of HNF4α targets in xenograft of MHCC-97H cells expressing WT or S304D mutant HNF4α. n = 11. ±SEM, Fisher LSD test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: AMPK CA mice were previously described ( ) (the Jackson Laboratory, #034797).

Techniques: Control, Western Blot, Binding Assay, Expressing, Mutagenesis, Gene Expression