Fig 1: TM6SF2 silencing affectsoxydative phopshorylation (OXPHOS)and when combined with MBOAT7 deletion switches up to metabolic reprogramming. (A) MT-COX1 levels were evaluated by Western blot and normalized to SDHA. (B) MT-COX1 protein expression was measured by enzyme-linked immunosorbent assay (λ = 600 nm) and normalized to SDHA levels (λ = 405 nm). (C) Complex I enzymatic activity was evaluated biochemically and normalized to citrate synthase (CS). (D) The total ATP rate was obtained by Seahorse XF Analyzers in live cells. (E and F) Biochemical measurements of lactate (mmol/L) and LDH (mmol/L) were assessed in cell supernatants and lysates, respectively. (G) Inverse correlation between secreted lactate levels and pH values. (H) The mRNA levels of Glucokinase (GCK), Phosphofructokinase (PFK), and Glyceraldehyde-3-phosphate dehydrogenase (G3PDH) were evaluated by reverse-transcription quantitative PCR and normalized to the β-actin housekeeping gene. At least 3 independent experiments were conducted. For the box plot, data are expressed as means and SE. Adjusted ∗P < .05 and ∗∗P < .01 vs Cas9+ and/or vs MBOAT7-/-.