VAP-A and VAP-B are not involved in LD homeostasis. (A) Western blot analysis of VAP proteins level in control HeLa cells (WT), HeLa cells transfected with control siRNAs (siCtrl), and with siRNAs targeting VAP-A (siVAP-A), VAP-B (siVAP-B), and both (siVAP-A + B). The band labeled with an * on VAP-B blot corresponds to cross-reactivity with VAP-A. (B) Representative confocal images of parental HeLa cells (WT) and HeLa cells transfected with control siRNAs (siCtrl), and with siRNAs targeting VAP-A (siVAP-A), VAP-B (siVAP-B), or both (siVAP-A + B). Cells were labeled with BODIPY 493/503 (LD, magenta) and Hoechst 33258 (nuclei, blue). The cell contour is delimited by a white dotted line. Scale bars: 10 µm. Images were acquired on a spinning-disk confocal microscope (Nikon CSU-X1, 100× NA 1.4). (C) Number (a) and area (b) of LDs in cells shown in B. Data are displayed as Superplots showing the mean number and area of LDs per cell (small dots), and the mean number and area of LDs per independent experiment (large dots). Independent experiments (n = 4) are color-coded. Means and error bars (SD) are shown as black bars. Data were collected from 98 (WT), 118 (siCtrl), 134 (siVAP-A), 129 (siVAP-B), and 135 (siVAP-A + VAP-B) cells. One-way ANOVA with Tukey’s multiple comparisons test (ns, not significant; n = 4 independent experiments). Source data are available for this figure: SourceData FS5.