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. 2013 Apr 8;288(20):14384–14390. doi: 10.1074/jbc.M112.441501

FIGURE 5.

FIGURE 5.

Cep57 affects Tektin 1 midbody localization. A and B, Western blotting performed after 60 h of control and Cep57 siRNA transfection. Cep57 (A) and Tektin 1 (B) in HeLa cells were detected by immunoblotting with the indicated antibodies. C, immunofluorescence images showing synchronized telophase HeLa cells treated with Cep57 or control siRNA stained for Tektin 1 (green), MKLP1 (red), and DNA (blue). Scale bar = 5 μm. D, immunostaining of MKLP1 (green), FLAG-Cep57 (green), and Tektin 1 (red) in HeLa cells transfected with Cep57 siRNA alone or together with FLAG-tagged siRNA-resistant Cep57 (siRes). DNA was labeled with DAPI (blue). Scale bar = 5 μm. E, immunostaining of Tektin 1 (red) in HeLa cells transfected with GFP-tagged Cep57 truncated mutants (green). Scale bar = 5 μm. F, immunofluorescence images of HeLa cells stained for α-tubulin (green). The cells were transfected with Tektin 1 or Cep57 siRNA and synchronized to telophase. DNA was labeled with DAPI (blue). Scale bar = 5 μm. G, percentage of cells without a visible Flemming body at 60 h after the indicated siRNA treatment (mean of three separate trials). A minimum of 200 cells were counted per sample. Error bars represent ± S.E. ***, p < 0.001; n.s., not statistically significant. siTektin1, Tektin 1 siRNA; siCep57, Cep57 siRNAs.