Fig. 6: ESCRTs are recruited at the MB of neural progenitors but are absent in Cep55-knockout fibroblasts. | Nature Communications

Fig. 6: ESCRTs are recruited at the MB of neural progenitors but are absent in Cep55-knockout fibroblasts.

From: Cep55 promotes cytokinesis of neural progenitors but is dispensable for most mammalian cell divisions

Fig. 6

a Quantification (mean ± SD) of MBs with Cep55 present or absent in the indicated cell types. n = 3 mice per cell type; 193, 237, and 214 cells quantified, respectively. b–d Immunofluorescence images of intercellular bridges in the indicated cell types stained for DNA (DAPI), α-tubulin, MKLP1, and Cep55. Boxed areas are magnified on the right. Scale bar in magnified regions, 2 µm. Punctate cytoplasmic signal is nonspecific staining. e Plots showing the fluorescence intensity along the intercellular bridge, in arbitrary units (AU), from TTFs as in c and d. Ten cells per genotype quantified. SEM are shown. f Quantification (mean ± SD) of MBs with Chmp2B present or absent in the indicated cell types. n = 3 mice per cell type; 90, 151, 133, 86, and 67 cells quantified. P-values for TTFs were calculated using two-way ANOVA, each mean compared with control Cep55+/+ (plastic), followed by Dunnett’s multiple comparisons test. For “Chmp2B present” category, P-values are: 0.2057, 0.0001, and 0.0004, respectively. g–k Immunofluorescence images of the indicated cell types undergoing abscission stained for DNA (DAPI), α-tubulin, MKLP1, and Chmp2B. Boxed areas are magnified below (g–i) or above (j, k). Scale bar, 2 µm. l Quantification of Chmp2B signal (mean ± SD) in TTFs grown on glass or poly-l-lysine (PLL)-coated glass coverslips as in h–k. For Cep55+/+ TTFs, only Chmp2B-positive cells were analyzed. n = 8, 9, 10, and 12 cells analyzed. P-value calculated using Student’s two-tailed unpaired t-test. m Time-lapse images of control and Cep55−/− TTFs expressing mChmp4B-EGFP and stained with Sir-tubulin to visualize the microtubules of the ICB. The white arrowheads indicate the site of abscission on the intercellular bridge. n Quantification of mChmp4B-EGFP at the intercellular bridge as in m. Individual pairwise comparisons were performed using χ2-test (one-sided). The omnibus χ2-test P-value is < 0.0001. n = 25, 26, 23, and 19 cells were imaged. Source data for a, e, f, l, and n are provided as a Source Data file.

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