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. 2021 Oct 20;15:713336. doi: 10.3389/fncel.2021.713336

Figure 2.

Figure 2

IL-33 protects glioma cells from the insult of DNA damaging drug, TMZ. (A) Scramble- and IL33KD-C6 cells were seeded onto a 6-well plate at the amount of 200 cells per well, and then grown in 10% serum containing medium for 5 days. The cultures were exposed to TMZ (0, 100, and 200 μM) for another day. Alternatively, MOCK- and IL33oe-C6 cells were maintained in 10% serum containing medium for 7 days, and treated with TMZ (0, 100, and 200 μM) for another day. The colonies were quantified using NIH Imaging J. Data are shown as mean ± SEM from the three cell passages (n = 3). *P < 0.05, **P < 0.01 vs. Vehicle. (B) MOCK- and IL33oe-C6 cells were treated without or with TMZ at 200 μM for 48 and 72 h. Total proteins collected from the cultures were subjected to Western Blot assay to examine the levels of γH2AX, IL-33, and GAPDH. The γH2AX protein levels in each immunoblot were normalized by the level of GAPDH (a loading control). The results are presented as mean ± SEM from three cell passages (n = 3). **P < 0.01, ***P < 0.001 vs. the relative control culture; ##P < 0.01, ###P < 0.001 vs. MOCK. (C) The expression of Rad51 and BRCA1 mRNA in MOCK- and IL33oe-C6 were measured by qPCR analysis. The results are presented as mean ± SEM from the five cell passages (n = 5). *P < 0.05, **P < 0.01 vs. MOCK. TMZ, temozolomide; qPCR, quantitative PCR.