Rev-erbα circadian power fraction does not correlate with nuclear MAL. (A) Confocal microscopy images of cells under different conditions (high-density control, low-density control, micropatterned cells of 1,600 and 900 µm2, low-density treated for 24 h with jasplakinolide 1 µM, latrunculin A 200 nM, cytochalasin D 1 µM, para-nitro-blebbistatin 10 µM, and low-density cells grown on polyacrylamide gels with a stiffness of 300 Pa). The cells were stained with an anti-MAL antibody (green, top) and Hoechst (blue, bottom); the cell perimeter is represented with a dashed white line. Scale bar, 20 µm. (B) Violin plots representing the distribution of the single-cell MAL nuclear to cytosolic ratios of the conditions depicted in A; n = 215, 254, 208, 332, 163, 140, 165, 157, and 162, respectively, from three to four experiments depending on the condition. Medians and interquartile ranges are depicted as white circles and black bars, respectively. The P values of the comparisons of every condition against the low-density, obtained with a two-sided Wilcoxon rank sum test, are represented in Table S1. (C) Dependence of circadian power fraction with MAL nuclear to cytosolic ratio of all the aforementioned conditions. The values represented are the means of the medians of each independent experiment for every condition. The error bars refer to the corresponding SD. Pearson’s correlation coefficient is r = −0.19 with a P value of 0.63.
