Fig. 6: POC1A and POC1B act together in centriole biogenesis. | Nature Communications

Fig. 6: POC1A and POC1B act together in centriole biogenesis.

From: An interaction network of inner centriole proteins organised by POC1A-POC1B heterodimer crosslinks ensures centriolar integrity

Fig. 6

a EM images of G1 centrioles from POC1A−/−, POC1B−/− and POC5−/− cells. The knockout cells lines have broken centrioles (as seen in the longitudinal view, magenta arrows) with the most proximal region usually intact (indigo arrow). This phenotype can be also observed by U-ExM. Cross-sections of these centrioles from proximal and distal regions show loss of entire MT triplets (magenta arrows) and in the case of POC1A−/− and POC5−/− deformation or loss of the inner scaffold structure in the distal half of the centrioles. Scale bars: 200 nm (EM) and 100 nm (U-ExM). b Quantification of longitudinal centrioles from (a). POC1B−/− centrioles show already defects at around 150 nm, whereas POC1A−/− and POC5−/− centrioles have mostly defects at 190-200 nm. Data are presented as mean ± SD. Statistics were derived from two-tail unpaired t-test. n = 20 (Control), 14 (POC1A−/−), 6 (POC1B−/−), 19 (POC5−/−). c IF images of interphase POC1A/B−/− double knockout cells show loss of centrosomal signal. Green: γ-tubulin, red: PCNT. Scale bars: 5 µm, magnification scale bars: 1 µm. d Percentage of the cells from (c) showing co-localisation of γ-tubulin and PCNT. Data are presented as mean ± SD. N = 3 biologically independent experiments, n > 100 cells per cell line for each experiment. e Control and POC1A/B−/− cells were stained against CEP44 (green) and PCNT (red) to detect the proximal part of centrioles. Scale bars: 5 µm, and 1 µm for the inset magnifications. f Percentage of cells from (e) showing CEP44 and PCNT co-localisation. Data are presented as mean ± SD. N = 2 biologically independent experiments, n > 100 cells per cell line for each experiment. g Mitotic spindle configurations observed in control and knockout cell lines. Green: α-tubulin, red: γ-tubulin, magenta: CDK5RAP2. Scale bars: 5 µm. h Percentage of cells in the knockout cell lines showing indicated spindle configuration in g. Data are presented as mean ± SD. N = 2 biologically independent experiments, n > 50 cells per cell line for each experiment. Source data are provided as a Source Data file.

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