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. 2011 Jan 18;124(4):532–539. doi: 10.1242/jcs.079731

Fig. 5.

Fig. 5.

Knockdown of Chfr in Stil-depleted cells restores growth rate and spindle bipolarity. (A) Immunoblot confirming Chfr knockdown in 1022M cells expressing shRNA targeting Chfr. Cbl was used as a loading control. (B) Increased growth rate of 1022M cells after shRNA knockdown of Chfr (from a doubling time of 72 hours to 30 hours). t-test: *, P<0.003, **, P<0.05. Error bars indicate s.e. (C) Immunostaining for α-tubulin (green) and γ-tubulin (red) in 1022M cells expressing shRNA targeting Chfr demonstrates partial correction of the abnormal centrosomal phenotypes caused by absence of Stil (centrosomes are marked by white arrows). (D) Immunoblotting of 3T3 cells (polyclonal populations) stably expressing shRNA targeting Stil, Chfr or both. (E) Immunostaining for α-tubulin (green) and γ-tubulin (red) shows restoration of bipolar centrosomal phenotypes in 3T3 cells with knockdown of Stil and Chfr. (F,G) Analysis of mitotic index (F) and growth rate (G) demonstrates that knockdown of Chfr partially corrects the defects caused by knockdown of Stil in these 3T3 cells. Error bars indicate s.e. Scale bars: 11 μM.