Fig. 2: ch-TOG transiently localizes to the PCM.
From: Microtubule nucleation and γTuRC centrosome localization in interphase cells require ch-TOG
a Maximum intensity projections of 3D-SIM images of centrosomes stained for endogenous or recombinant ch-TOG (ch-TOG-GFP), in the presence or absence of microtubules. Cells were costained with antibodies against acetylated α-tubulin or antibodies against γ-tubulin and NIN as indicated. b Total centriolar intensities of endogenous ch-TOG staining in cells with or without microtubules as in (a) were quantified, normalized to the average intensity in cells with microtubules, and plotted. N = 3 independent experiments, total number of cells analyzed per condition: 141 (MTs + ) and 144 (MTs -), respectively. **p = 0.0018. The horizontal bars and whiskers indicate median and interquartile range, respectively, of the plotted data points. Statistical significance was determined by unpaired, two-tailed t test with Welch’s correction. c Intensities of endogenous ch-TOG staining at the PCM and at subdistal appendages in cells with or without microtubules as in (a) were quantified, normalized to the average intensities in cells with microtubules, and plotted. N = 3 independent experiments. Total number of cells analyzed for PCM staining: 55 (MTs+) and 56 (MTs−). *p = 0.0368. Total number of cells analyzed for subdistal appendage staining: 55(MTs+) and 56 (MTs−). *p = 0.0142. The horizontal bars and whiskers indicate median and interquartile range, respectively, of the plotted data points. Statistical significance was determined by unpaired, two-tailed t test with Welch’s correction. d Cell lysates prepared from U2OS cells with and without microtubules were analyzed by immunoblot using antibodies against the indicated proteins. Observed in two independent experiments. e Microtubules in U2OS cells were depolymerized, allowed to re-grow for the indicated time points, fixed and stained with antibodies against the indicated proteins. Illustrations indicate centriole orientations in the respective images. Performed twice with similar result. Scale bar, 1 μm. Source data are provided as a Source Data file.
