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. 2023 May 24;13:8435. doi: 10.1038/s41598-023-35712-5

Figure 2.

Figure 2

The effects of P2X7 receptor (P2X7R) antagonism by AZ10606120 (AZ) compared to conventional temozolomide (TMZ) chemotherapy on the U251 human glioblastoma cell line. U251 human glioblastoma cells were cultured and treated at 80% confluency with 15 μM AZ, 50 μM TMZ and a combination of AZ + TMZ for 72 h. The control group were untreated cells. U251 cultures were counterstained with DAPI for 1 h at room temperature. Images were acquired with a Nikon Ti-E inverted fluorescence motorised microscope equipped with a sCMOS Andor Zyla camera at 20 × objective. The total number of DAPI + cells were quantified across 16 random fields for each sample and treatment group. (A) Comparison of AZ 15 μM with TMZ in U251 cells. Results were yielded from a one-way ANOVA with Tukey’s HSD post-hoc, F(2, 33) = 64.87, p < 0.0001. N = 12 replicates. Untreated versus TMZ: mean difference = 578.3 cells, 95% CI = 259.3 to 897.4; Untreated versus AZ 15 μM: mean difference = 1470 cells, 95% CI = 1151 to 1789; AZ 15 μM versus TMZ: mean difference = − 891.5, 95% CI = − 1211 to − 572.5. ***p = 0.0003, ****p < 0.0001. Error bars represent SEM. (B) Comparison of 15 μM AZ and AZ-TMZ co-therapy with TMZ in U251 cells. Results were yielded from a one-way ANOVA with Tukey’s HSD post-hoc. N = 12 replicates, F(3, 44) = 40.77, p < 0.0001. Untreated versus AZ 15 μM: mean difference = 3435 cells, 95% CI = 2035 to 4835; Untreated versus TMZ: mean difference = − 1177 cells, 95% CI = − 2577 to 223.2, p = 0.13; Untreated versus AZ + TMZ: mean difference = 3419 cells, 95% CI = 2019 to 4819; AZ 15 μM versus TMZ: mean difference = − 4612 cells, 95% CI = − 6012 to − 3212. AZ 15 μM versus AZ + TMZ: mean difference = − 16.33 cells, 95% CI = − 1416 to 1384, p > 0.99; TMZ versus AZ + TMZ: mean difference = 4596 cells, 95% CI = 3196 to 5996. *p < 0.0001. Error bars represent SEM. (C) Image representation of U251 cells treated with 15 μM AZ, TMZ, AZ + TMZ and control.