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. 2022 Feb 18;13:815308. doi: 10.3389/fphar.2022.815308

FIGURE 2.

FIGURE 2

GDC potentiates paclitaxel-induced cytotoxicity in NSCLC cells. A549 cells were preincubated with DMSO (vehicle; Veh) or GDC-0449 (GDC; 20 μM) for 1 h followed by treatment with various doses of PTX for 24 h. Cell viability was determined by trypan blue exclusion (A) and MTS/PMS activity (B) assays. (C) Cells were preincubated with Veh or GDC (20 μM) for 1 h followed by PTX (20 nM) treatment for 24 h. Apoptotic cells were identified by morphology under light microscopy (×20 objective). G + P, GDC + PTX. (D) Cells were treated with GDC (20 μM) and PTX (20 nM) for 24 h, with or without preincubation with the pan-caspase inhibitor, Z-VAD-FMK (Z-VAD; 100 μM). G + P, GDC + PTX; G + P + Z-VAD, GDC + PTX + Z-VAD. Cell viability was analyzed by the MTS/PMS activity assay. (E) MRC-5 cells were preincubated with Veh or GDC (20 μM) for 1 h followed by PTX (20 nM) treatment for 24 h. Cell viability was assessed with the MTS/PMS assay. (F) A549 cells were preincubated with GDC (20 μM) for 1 h followed by PTX (20 nM) treatment for an additional 1 h. Whole cell lysates were collected and immunoblotted with anti-GLI1, anti-Shh, and anti-β-actin antibodies. Protein band intensities were analyzed using ImageJ. G + P, GDC + PTX. *: p < .05; indicates a statistically significant difference between indicated groups. All experiments were performed at least three times. n = 3 for all groups.