Centrosome position is affected by lowering the levels of LCBs. (a) Representative immunofluorescence images of RPE-1 cells upon knockdown of scramble (Control), SPTLC1, or SPTLC2 cultured for 96 h, or treated with 5 µM myriocin or 10 µM fumonisin B1 in lipid-depleted medium for 48 h. Green, anti-alpha-tubulin; purple, Hoechst 33342 (DNA); and red, anti-lamin B1. Scale bar, 5 µm. (b) Quantification of yH2AX in RPE-1 cells upon knockdown of scramble (Control), SPTLC1, or SPTLC2. N = 5 western blots, NS, P > 5 e-2, one-way ANOVA. (c) Representative immunofluorescence images of RPE-1 cells with BRCA1 deletion (left) showing increased 53BP1 foci (red). RPE-1 cells cultured in LD-FBS (middle) or arrested in G2 in LD-FBS with abnormal nuclear morphology (right) do not show 53BP1 foci. Scale bar, 5 µm. (d) Quantification of cells shown 53BP1 foci (n > 200). (e) Representative immunofluorescence images of RPE-1 cells upon knockdown of scramble (Control), SPTLC1, or SPTLC2 cultured for 96 h. Scale bar, 10 µm. (f) Quantification of the centrosome distance to the NE in RPE-1 cells. *P < 1 e-4, one-way ANOVA. (g) Representative immunofluorescence images of RPE-1 cells treated with 5 µM myriocin, 40 µM C75, 10 µM TOFA, 10 µM cerulenin, or 1 µM lovastatin cells for 24 h. Scale bar, 10 µm. (h) Quantification of the centrosome distance to the NE in RPE-1 cells. *P < 1e-4, one-way ANOVA. NS, P > 0.05.
