Figure 5: Knockdown of Rb and FZR1 overcomes cell cycle arrest induced by CDK4/6-inhibitor treatment of human breast cancer cell lines.
From: Rb and FZR1/Cdh1 determine CDK4/6-cyclin D requirement in C. elegans and human cancer cells
(a–d) Representative immunofluorescence images of a field of CAMA-1 (a,b) and T47D (c,d) cells, following visualization of DNA replication by EdU incorporation and staining, and nuclei with DAPI staining. Cells were first transfected with nonspecific control siRNAs (siCTRL) (a,c), or combined FZR1- and Rb-specific siRNAs (b,d), treated with the CDK4/6 inhibitor PD-0332991 (500 nM) and subsequently incubated with EdU. Scale bar, 10 μm. (e,f) The percentage of EdU-positive nuclei counted for CAMA-1 (e) or T47D (f) cells treated with siRNA pools as indicated, in the absence or presence of PD-0332991 (500 nM). (g–h) Western blottings of CAMA-1 (g) or T47D (h) total cell lysates, following transfection with the indicated siRNA pools. Blots were probed with either anti-Rb or anti-FZR1 antibodies. All aspects of the experiments were performed twice, with similar results.
