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. 2016 Jun 23;113(28):7828–7833. doi: 10.1073/pnas.1607421113

Fig. S4.

Fig. S4.

Expression and localization of siRNA-resistant GFP-Mps1 constructs. (A) HeLa cells were transfected with either siControl (siRNA against Lamin A/C) or siMps1 for 24 h before being transfected with the indicated siRNA-resistant construct for 48 h. For the final 24 h, cells were arrested in S-phase by HU treatment. Cells were lysed, separated via SDS/PAGE, and blotted with indicated antibodies. Total transfected protein amount was assayed by GFP immunoblot; knockdown of endogenous Mps1 was confirmed by Mps1 (N1) immunoblot, and alpha tubulin (α-tub) served as a loading control. (B) Percentage of S-phase–labeled (by BrdU-incorporation) GFP+ cells with GFP signal colocalizing with γ-tubulin at centrosomes, presented as the mean ± SD of n = 3 independent experiments of n = 100 cells for each experiment.