Fig. 5.

FOCAD promotes mitochondrial function via enhancing the activity of FAK. FOCAD-knockout (FOCAD-KO) cell line was established by CRISPR/CAS9 in A549 cells, and the effect of FOCAD on FAK activity was evaluated by western blot (A). The relative quantification of immunoblot results was showed in 5B. FOCAD overexpression was induced by lentivirus transduction in A549 cells, and both FOCAD-wild type (FOCAD-WT) and FOCAD-overexpressed (FOCAD-OE) cells were treated with FAK inhibitor, PF-573228 (3 μM for 24 h). Then the cells were harvested for western blot detection (C–D). Besides, the function of PF-573228 on mitochondrial TCA cycle (E–F) and Complex I activity in mitochondrial ETC (G and Fig. S6) were also evaluated respectively. The ferroptosis model was induced by erastin (5 μM for 24 h) treatment, and the cell viability (H), MDA (I) and 4-HNE (J), as well as the expression of PTGS2 (K) were measured respectively to evaluate ferroptosis in different groups. Results were expressed as mean ± SD (n = 3), and the P value less than 0.05 was considered statistically significant. In Fig. 5A–D and 5G, #: P < 0.05 compared with FOCAD-WT/FOCAD-WT + DMSO group. In Fig. 5E–F and 5H–5K, *: P < 0.05 compared with the FOCAD-WT group in the first condition. #: P < 0.05 compared between FOCAD-WT group and FOCAD-OE group in same condition. &: P < 0.05 compared with the FOCAD-OE group in the first condition.