Fig. 5: CDK4/6 and KRAS inhibitor commonly target many pathways to suppress PDAC.
From: Deprivation of EGFR signal causes senolysis in PDAC with CDK4/6 inhibition
A IB of the indicated proteins in MIA PaCa-2 cells treated with or without 1 µM sotorasib or 10 µM palbociclib for 0-72 h. B Pull down assay of GTP-KRAS in MIA PaCa-2 cells treated with or without 1 µM sotorasib for 0-72 h. C, D Enrichment plots of HALLMARK_Apoptosis, HALLMARK_DNA repair, and HALLMARK_IL6_JAK_STAT3_signaling pathway from GSEA (MIA PaCa-2 cells). E RT-qPCR determination of IL-6 in MIA PaCa-2 cells treated with or without 1 µM sotorasib for 0-72 h. F Enrichment plots of EGFR_UP.V1_UP pathway from GSEA (MIA PaCa-2 cells). G, H RT-qPCR determination of EGF family of ligands and Sprouty family members in MIA PaCa-2 cells treated with or without 1 µM sotorasib for 0-72 h. I An enhanced volcano plot was created based on total fraction RNA-seq data [sotorasib (+) vs palbociclib (+)] using all differentially expressed genes (DEGs) (17,622 genes). Y-axis shows the mean expression value of log10 (p-value), and the x-axis displays the log2 fold change value. Red points: up-regulated DEGs; Blue points: down-regulated DEGs. J Representative images of SA-β-gal staining in MIA PaCa-2 cells simultaneously treated with or without 1 µM sotorasib and 10 μM palbociclib for 72 h. Quantitation of SA-β-gal positive cells from 3 or more randomly chosen fields. K Representative flow cytometry profiles of annexin V/PI double staining in MIA PaCa-2 cells treated as in (J). Quantitation of apoptotic cells (% = early apoptotic cells in Q2 + late apoptotic cells in Q3). α-tubulin was used as a loading control. DMSO was used as a vehicle. All data are presented as mean ± SD of three independent experiments. One-way ANOVA followed by Tukey’s post-hoc test was performed in (G, H, J and K). *p < 0.05, **p < 0.01, ***p < 0.001.
