Fig. 2: RAD18 promotes TMZ tolerance in Astrocyte and GBM cell lines. | Nature Communications

Fig. 2: RAD18 promotes TMZ tolerance in Astrocyte and GBM cell lines.

From: Trans-lesion synthesis and mismatch repair pathway crosstalk defines chemoresistance and hypermutation mechanisms in glioblastoma

Fig. 2

a Representative immunoblot showing the effect of 100 µM TMZ treatment (24 h) on PCNA mono-ubiquitination in NHA and NHARas cells transfected with siCon or siRAD18. b Effect of RAD18 knockdown on TMZ sensitivity in NHA and NHARas cells. (**p = 0.002, *p = 0.0174). Clonogenic survival assays showing TMZ-sensitivities of parental (RAD18 +/+) and RAD18 −/− U373 (c, ***p = 0.0006) and U87 cells (d, ***p = 0.0009). Clonogenic survival assays showing TMZ-sensitivities of RAD18 +/+ and RAD18 −/− LN18 cells treated with or without 20 µM O6BG (e, *p = 0.0128) or 5 mM MeOX (f, *p = 0.0111). g Clonogenic survival assay TMZ-sensitivities of RAD18 +/+ and RAD18 −/− U373 cells treated with (**p = 0.004) or without (***p = 1.0460e-05)10 mM MeOX. h, i TMZ sensitivity of RAD18 +/+ and RAD18 −/− U373 cells cultured on viable organo-type brain slice explants. The middle panel shows representative bioluminescence images of firefly luciferase-expressing U373 RAD18 +/+ and RAD18 −/− cells cultured on brain slices and treated with indicated concentrations of TMZ for 4 days. To measure TMZ sensitivity, bioluminescence on day 4 post-TMZ treatment was normalized to day 0 for each treatment group (n = 4). 100 µM TMZ ***p = 0.0003, 250 µM TMZ ***p = 9.4745e-05. Data points represent mean ± SD of triplicate determinations; P values were determined by two-sided t test (b–g, i). Experiment of (a–g) and (I) had been repeated at least three times with similar results. Source data are provided as a Source Data file. (h) was created with BioRender.com.

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