Figure 2. TBK1-deficient hepatocytes exhibit increased susceptibility to mitochondrial stress.

(A) Mitochondrial ROS levels measured by MitoSOX staining in WT and TBK1 KO HepG2 cells (n = 4). (B) Mitochondrial ROS levels in primary hepatocytes transfected with siNC or siTbk1 (n = 3). (C, D) mRNA expression of proinflammatory genes Saa3 (C) and Il1b (D) in siNC or siTbk1-transfected primary hepatocytes (n = 3). (E, F) Cell viability measured by CCK-8 assay in primary hepatocytes under basal conditions (E) and following 10 mM of CCCP treatment (F) (n = 3). (G) Bax/Bcl2 mRNA expression ratio in primary hepatocytes under CCCP-induced stress (n = 3). (H) Immunoblot of cleaved caspase-3 in primary hepatocytes treated with CCCP. (I) Basal ¹⁴C-palmitate oxidation activity in primary hepatocytes from flox and LTKO mice (n = 6). (J) Fold change in CCCP-induced suppression of palmitate oxidation activity in flox versus LTKO hepatocytes (n = 6). (K) Immunoblot analysis of phosphorylated and total STAT3 and p38 in liver tissues from flox and LTKO mice (n = 6). (L,M) Quantification of phospho-STAT3 (L) and phospho-p38 (M), normalized to their respective total protein levels (n = 6). * p < 0.05, **p < 0.01, ***p < 0.001, ***p < 0.0001. Data are presented as mean ± SD (A–G, I, J) or mean ± SEM (L, M). Statistical significance was assessed by unpaired two-tailed Student’s t-test (E, F, J, L, M) or two-way ANOVA (A–D, G, I).