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. Author manuscript; available in PMC: 2018 Sep 1.
Published in final edited form as: Mol Cancer Ther. 2017 Jun 15;16(9):1751–1764. doi: 10.1158/1535-7163.MCT-17-0040

Figure 3. Rb knockdown induces palbociclib resistance.

Figure 3

(A) Western blot showing successful knockdown of Rb and indicated proteins in SK-LMS1 cells expressing non-targeting control (NTC) shRNA or Rb shRNA. Rb is significantly depleted, while other drug targets are not significantly altered. Actin serves as a loading control on the gels. (B) Cell number after indicated number of days and doubling time (ns = not significant compared to NTC). (C) IC50 for 4, 6 and 8 days of palbociclib treatment in SK-LMS1 cells, using MTT as a readout of survival at day 12 (n=3), *p < 0.05, **p < 0.01. (D) Cell number after 6 days of palbociclib treatment with the indicated doses, normalized to vehicle in each cell line (DMSO) (n=3), *p < 0.05 (NTC vs KD1+KD2), **p < 0.01 (NTC vs KD1+KD2). (E) Cell cycle change in G1 and S phase for cells treated with palbociclib for 6 days prior to fixation and flow cytometry analysis. (F) Sensitivity to AZD1775 treatment for 24 or 48 hours using MTT as a readout of survival at day 12/13 (n=2). Right panel shows IC50 values derived from the individual experiments, ns = not significant.