Fig. 2: Pharmacological inhibition of pBADS99 in PDAC cells induces G0/G1 cell cycle arrest and promotes apoptotic cell death. | Cell Death & Disease

Fig. 2: Pharmacological inhibition of pBADS99 in PDAC cells induces G0/G1 cell cycle arrest and promotes apoptotic cell death.

From: Concurrent inhibition of pBADS99 synergistically improves MEK inhibitor efficacy in KRASG12D-mutant pancreatic ductal adenocarcinoma

Fig. 2: Pharmacological inhibition of pBADS99 in PDAC cells induces G0/G1 cell cycle arrest and promotes apoptotic cell death.

A Western blot analysis was used to assess the level of BAD proteins in KRAS-mutant PDAC cells (Panc-1, SW1990, AsPC-1, Capan-1, CFPac-1 and Capan-2). β-ACTIN was used as input control. The sizes of detected protein bands in kDa are shown on the left. The densitometric analysis of protein blots is shown below along with the mutational status of the PDAC cell lines. Oncogene (red): KRAS; Tumor suppressor genes (black): TP53, CDKN2A, SMAD4 and BRCA2. B Western blot analysis was used to assess the level of BAD protein in PDAC cells (SW1990, Panc-1, AsPC-1, and BxPC-3) after treatment with 0-10 μM of NCK for 72 h. β-ACTIN was used as input control. The sizes of detected protein bands in kDa are shown on the left. C Crystal violet staining of foci in colonies of PDAC cells after exposure to 0-10 μM of NCK for 9 days. D Microscopic visualization of Calcein-AM (green) stained colonies (live) and BOBO-3 Iodide (red) stained cell debris (dead) generated by PDAC cells cultured in 3D Matrigel after exposure to 0-10 μM of NCK for 12 days. Scale bars, 100 μm. BF: Bright-field image; Merge: Merged image of Live and Dead. E. Flow cytometry analysis of PI staining for cell cycle state of PDAC cells measured after treatment with 0-10 μM of NCK for 48 h using flow cytometry analysis as described in materials and methods. Data represent means ± SD (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001. F. Flow cytometry analysis of Annexin-V and propidium iodide (PI) staining of apoptotic cell death of PDAC cells measured after treatment with 0-10 μM of NCK for 72 h using flow cytometry analysis as described in materials and methods. The upper left quadrant (Annexin V−, PI + ) represents cell debris, the upper right quadrant (Annexin V + , PI + ) represents late apoptosis, the lower right quadrant (Annexin V + , PI-) represents the early apoptosis and the lower left quadrant (Annexin V-, PI-) represented live cells. Data represent means ± SD (n = 3). *P < 0.05, **P < 0.01, and ***P < 0.001.

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