(
A) Representative fluorescence images of control HeLa cells treated with 2 µM latrunculin B (LatB) or not (untreated) that were fixed and stained with Alexa Fluor 546-phalloidin. Image of untreated cells was deconvolved using the blind-algorithm package of LAS X. (
B) Comparison of cFRET in isotonic and hypotonic buffer of HeLa cells expressing A-CFP/E-YFP and treated with latrunculin B (LatB; n = 8 dishes with 26 cells) as in (
A) or methyl-β-cyclodextrin (MbCD; n = 10 dishes with 46 cells) as in (
C) with untreated cells (data from
Figure 1E). Data represent individual cells (diamonds) and mean ± s.e.m. (
C) Left: Representative fluorescence images of control HeLa cells treated with 5 mM methyl-β-cyclodextrin (MbCD) or not (untreated) that were fixed and stained with filipin. Right: filipin fluorescence intensity of untreated and MbCD-treated HeLa cells. n = 5 with over 300 cells per condition, data represent individual FOVs (diamonds) and mean ± s.e.m. Scale bars, 20 µm. Statistics: *p<0.05; **p<0.005; n.s., not significant, Student’s
t-test.