Figure 6. TAZ-induced Cybb/NOX2 contributes to the development of NASH-HCC tumors.

(A-G) AAV8-TBG-Cre-treated Rosa^NICD mice were fed the NASH diet and, 2 months later, injected with AAV8-H1-Scr or AAV8-H1-shCybb. The mice were analyzed at month 4. (A) Experimental scheme. (B) Nox2, γH2AX, and H2AX immunoblots from surrounding tissue (ST), with quantification (n = 3; means ± SEM; ***p<0.001, ****p<0.0001 by two-way ANOVA/Sidak’s post-hoc analysis). (C) Livers (arrows, tumors) and tumor numbers/mouse. (D) Liver sections were stained with H&E (upper images) and Sirius red (lower images) and quantified for the number of inflammatory cells and the percent Sirius red-positive area. Bars, 200 μm. (E) Percent TUNEL⁺ cells in non-tumor areas. (F) Plasma ALT. (G) Average tumor diameter and percent Ki67⁺HNF4α⁺ cells in non-tumor areas. For C-G, n = 6–8 mice/group; means ± SEM; *p < 0.05 by Student’s t-test. (H-N) AAV8-TBG-Cre-treated Rosa^NICD mice were fed the NASH diet and, 2 months later, injected with AAV8-H1-shTaz and either AAV8-TBG-LacZ or AAV8-TBG-Cybb. The mice were analyzed at month 4. (H) Experimental scheme. (I) Nox2, γH2AX, and H2AX immunoblots from ST, with quantification (n = 4; means ± SEM; **p<0.01, ***p<0.001 by two-way ANOVA/Sidak’s post-hoc analysis). (J) Livers (arrows, umors) and tumor numbers/mouse. (K) Liver sections were stained with H&E (upper images) and Sirius red (lower images), with quantification of inflammatory cells and percent Sirius red-positive area. Bars, 200 μm. (L) Percent TUNEL⁺ cells in non-tumor areas. (M) Plasma ALT. (N) Average tumor diameter and percent Ki67⁺HNF4α⁺ cells. For J-N, n = 7 mice/group; means ± SEM; *p < 0.05 by Student’s t-test.