Figure 1.

(a) Epididydmal fat pad weight after feeding C57BL6/J mice either chow or HFD for 2 weeks (n = 6–10 animals/group). (b) Adipose cell size distribution obtained by coulter counter analysis of epididydmal fat tissue collected from mice in a). Data displayed as average of n = 4–6 samples/group. The fraction of cells to the right of the dashed line is defined as the large cells, in line with previous studies⁸. (c) Representative confocal images of isolated adipocytes (from chow or HFD-fed mice) stained with BODIPY (green) to show the neutral lipid droplet (top panel), and phalloidin (black) to visualize F-actin (bottom panel). (d) Representative TIRF images of isolated adipocytes from HFD-fed mice stained for different cytoskeletal markers (Vimentin, Tubulin, Actin (phalloidin)). (e) ImageJ plugin ridge detection was used to trace actin filaments (detected with phalloidin stain) in images collected with confocal and TIRF microscopy. (f) Quantification of traces detected in (e), data presented as Trace Area/ROI (px²), n = 40 ROI/condition. (g) Graph illustrating correlation between actin filaments and adipose cell size after 2 weeks of either chow (white squares) or HFD (black squares) feeding. Cell size was determined simultaneously while collecting images of phalloidin stain. (h) same as in (g), also including data from 4 weeks of either chow or HFD. Data in (a,f) presented as mean ± SD, ****p ≤ 0.0001. Scale bar = 20 µm.