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. Author manuscript; available in PMC: 2021 Sep 1.
Published in final edited form as: Cancer Res. 2020 Dec 15;81(5):1347–1360. doi: 10.1158/0008-5472.CAN-20-2275

Figure 4: Pharmacological induction of p27 in PDAC models.

Figure 4:

(A) Growth of 1222 and 226 cells following SKP2 knockdown. Bars represent mean and SD (2-way ANOVA). (B) Western blotting on 1222 and 226 cells following SKP2 knockdown. (C) Growth of 1222 and 226 cells that were treated with palbociclib (250 nM) in combination with SKP2 inhibitor, pevonedistat (500 nM). Graphs represent mean and SD (2-way ANOVA). (D) Western blotting on 1222 and 226 cells following combination treatment with palbociclib and pevonedistat. (E) Heat map showing the changes in relative growth rate of 1222 cells to a panel of targeted therapies in combination with DMSO and palbociclib (200 nM). (F) Growth of 1222 cells treated with palbociclib (200 nM) in combination with different MEK inhibitors, trametinib (Tram) (20 nM), pimasertib (500 nM) and PD318088 (500 nM). Graph represents mean and SD (2-way ANOVA). (G) Western blot on 226 cells following kRAS knockdown. (H) Western blot on 1222 cells following the combination treatment with palbociclib (200 nM) and trametinib (20 nM) and pimasertib (Pima) (500 nM). (I) Immunoprecipitation of cyclin D1 from 1222 cells treated with palbociclib (200 nM) +/− trametinib (20 nM) up to 48 H. Coimmunoprecipitated P27 and CDK4 were determined by immunoblotting. (J) In vitro CDK4 kinase assay associated on 1222 cells treated with palbociclib (200 nM) +/− trametinib (20 nM) up to 48 H. (K) Immunoprecipitation of P27 from 1222 cells treated with palbociclib (200 nM) +/− trametinib (20 nM) up to 48 H. Coimmunoprecipitated P27 and CDK2 were determined by immunoblotting. (L) Representative images of 1222 cells stably expressing the CDK2 sensor following 48 H exposure with palbociclib (200 nM) +/− trametinib (20 nM). Column graph indicates ratio of number of cells with cytoplasmic localization to number of cells with nuclear localization of the HDHB-mCHERYY protein at the indicated conditions. Bars represent the mean and SD (t-test). Graphs represent 2 independent experiments with 3 replicates. (*p< 0.05, **p<0.01, ***p<0.001).