Extended Data Fig. 4: Combined FDX1 deletion and FDX2 depletion elicits severe defects in growth and mitochondrial Fe/S proteins.
From: Functional spectrum and specificity of mitochondrial ferredoxins FDX1 and FDX2
a Cumulative growth of HEK293 cells from Fig. 2f was calculated from cell counts at the three harvests on day 3, (n = 6 each), day 6 (n = 5, Control(CC)/FDX2-si; n = 6, FDX1-CC2/FDX2-si) and day 9 (n = 2, Control(CC)/FDX2-si; n = 4, FDX1-CC2/FDX2-si) after the first electroporation. Values were presented relative to those of mock control cells (no CRISPR, no RNAi treatment; set to 100%, dashed line; mean ± SEM). b HEK293 cells were transfected with FDX1-directed gRNA-encoding plasmids (CC1 to CC3) and subsequently with FDX2-directed siRNAs similar to Fig. 2f. Cell samples were obtained at the specified time points and subjected to immunostaining of the indicated mitochondrial proteins or lipoyl cofactor. The observed molecular weights are given in parentheses. C-I, C-II, C-III, respiratory complexes I, II, and III. C-V, F1Fo ATP synthase. The immunoblot signals from cell samples with combined FDX1-CC2 and FDX2-si deficiency are representative for at least three independent experiments, and are here presented conjointly with FDX1-CC1 / FDX2-si and FDX1-CC1 / FDX2-si treated cells, respectively.