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. 2017 Jun 14;40(6):401–409. doi: 10.14348/molcells.2017.0032

Fig. 4.

Fig. 4

BFA inhibits the expression of Plk1 mRNA.

(A) BFA does not work in the ubiquitination of Plk1. HEK293T cells were co-transfected with HA-Ub and Flag-Plk1. The DMSO control or BFA was applied for 18 h. To prevent protein degradation, MG132 was applied to cells 3 h prior to cell harvest. Flag-Plk1 was immunoprecipitated with anti-Flag antibody and precipitates were subjected to immunoblotting analysis with the indicated antibodies. (B, C) BFA reduces the expression of Plk1 mRNA. hTERT-RPE cells were treated with BFA as indicated in Fig. 2. Total RNAs were subjected to semi-quantitative conventional RT-PCR analysis (B) and quantitative real-time RT-PCR analysis (C). (D) Overexpression of Plk1T210D and Dvl23E rescues BFA-induced defect of primary cilia disassembly. The schedule of transfection and serum-restimulation assay is drawn (upper). Data are mean ± SD from three independent experiments. Statistical analysis: ***P < 0.001, **P < 0.01 (unpaired two-tailed t-test). > 300 cells were counted for each sample (D).