Fig 1: TCHP interacts directly with PCM1 A, B(A) Immunoblot analysis showed HEK293 transfected for 48 h with expression vectors for TCHP, and FLAG-TCHP at N- and C-terminal. (B) Immunoblot analysis showed HEK293 transfected for 48 h with expression vectors for TCHP-FLAG C-terminal or constructs with deletions of the coiled-coil domain 1 (TCHP ?1) and 2 (TCHP ?2), as indicated in the scheme. For (A and B), total lysates were immunoprecipitated with anti-FLAG antibodies, and blots were probed sequentially with anti-PCM1, anti-FLAG and anti-ACTIN antibodies. IgG light chains are indicated with Red Ponceau staining. The input totals were analysed by parallel immunoblotting as a control for the level of expression.CAnti-PCM1 immunoprecipitation from HUVECs cells and TCHP and PCM1 immunoblot.DCo-localization of TCHP-V5 and centriolar satellite proteins. HUVECs were transduced with TCHP-V5 lentivirus and stained for anti-PCM1, anti-CEP290, anti-CEP72 and anti-V5 antibodies. Scale bars, 25 and 5 µm in the inset. Right panel: the two-channel intensity correlation of pixels corresponding to regions identified with TCHP-V5 and satellite markers (n = 90 cells; Pearson co-localization coefficient).ETCHP knock-down or control cells were stained for anti-PCM1 antibody. Panel below: quantification (n = 50 cells; unpaired t-test; **P = 0.004 vs. control). Scale bars, 50 and 5 µm in the inset.Data information: Statistical analyses were performed on at least three independent experiments. Data are represented as mean ± SD.