Fig. 3: Role of IL-32γ on inhibition of lung cancer stem cells.

a To study the potential role of IL-32γ against CSCs, CD133+ cells were cultured at 60–70% confluency and transfected with human IL-32γ vector as 1 µg/well of a six-well plate, after 0, 24, or 48 h of post transfection; cell proliferation assay was performed to investigate the efficacy of IL-32γ against CSCs. *p < 0.05. b CD133+ cells were transfected with human IL-32γ vector as 0, 5, 1, or 2 µg/well of a six-well plate, after 48 h of post transfection; cell proliferation assay was performed to investigate the efficacy of IL-32γ against CSCs. *p < 0.05. c Colony formation by control CD133+ and IL-32γ-CD133+ A549 cells. CD133 + isolated cells were cultured for 9 days in a six-well plate, which showed self-renewal capacity and tumor-sphere formation efficiency (after sphere formations, cells were stained with Crystal Violet) and photographed at 20× resolution. n = 5, *p < 0.05. d CD133+ cells were transfected with the IL-32γ vector at 2 µg or treated as mock/control for 48 h. Cellular DNA content was determined by flow cytometry, and the relative cell-cycle distribution is given in percentage. Total relative number of arrested cells at G2/M checkpoints have been plotted with a bar graph from three independent experiments. e CD133+ cells were transfected with human IL-32γ for 24 h, and then labeled with Annexin-V/PI. After that, apoptosis was analyzed by flow cytometry as described in the “Materials and methods” section. Data are representative of three independent experiments performed in triplicates. ***p < 0.001