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. 2022 Jan 20;9(8):2104055. doi: 10.1002/advs.202104055

Figure 5.

Figure 5

The binding of pyruvate to SSRP1 is required for tumor cell survival and irradiation resistance of glioblastoma. Data are representative of at least three independent experiments. A) SSRP1‐depleted U251 cells reconstituted with rSSRP1 WT or R54A were treated with etoposide (200 × 10–6 m) for indicated time. Cell viability was determined. Data represent the mean ± SD of the viability of the cells from three independent experiments (two‐tailed Student's t‐test). P values for comparisons between shSSRP1+rSSRP1 WT and shSSRP1+rSSRP1 R54A are shown. B) SSRP1‐depleted U251 cells reconstituted with rSSRP1 WT or R54A were supplemented with or without pyruvate (10 × 10–3 m, 3 h) and then treated with etoposide (200 × 10–6 m) for indicated time. Cell viability was determined. Data represent the mean ± SD of the viability of the cells from three independent experiments (two‐tailed Student's t‐test). C–E) U87/EGFRvIII cells stably expressing luciferase were infected with the lentivirus expressing shNT or shSSRP1 and the SSRP1‐depleted U87/EGFRvIII cells were reconstituted with the expression of rSSRP1 WT or R54A. These genetically modified cells (2 × 105 per mouse) were intracranially injected into randomized athymic nude mice (five mice per group) and then treated with or without IR (X‐ray) radiation (6 Gy). Bioluminescence imaging of tumor growth was carried out. C) Representative real‐time images were presented and D) the intensities of luciferase were quantified using living image software (PerkinElmer). Data represent the mean ± SD of luciferase intensity of five mice per group (two‐tailed Student's t‐test). E) Survival durations of these implanted mice were compared (Log‐rank test).