Figure 7
From: YB1 modulates the DNA damage response in medulloblastoma
YB1 depletion results in delayed radiation response in SHH and Group 3 Medulloblastoma (A1) 5.0e4 ONS-76 cells irradiated at 2.5Gy and 5Gy, harvested after 4 days, and counted (3.46e6 shSCR NT vs 3.02e6 shYB1 NT p = 0.081; 2.31e6 shSCR 2.5Gy vs 1.63e6 shYB1 2.5Gy p = 0.0026; 9.10e5 shSCR 5Gy vs 3.79e5 shYB1 5Gy p = 0.0251, n = 3). (A2) Doubling time calculated for ONS-76 (19.4 h shSCR NT vs 20.0 h shYB1 NT p = 0.9942; 21.7 h shScr 2.5Gy vs 24.1 h p = 0.8034; 29.1 h shScr 5Gy vs 42.5 h shYB1 5Gy p = 0.0015, n = 3). (A3) β-Gal stain of ONS-76 cells following radiation time course demonstrating increased senescence of YB1 depleted cells compared to irradiated control (5Gy shSCR vs 5Gy shYB1 95% CI = −22.64–(−7.91) p < 0.0001 two-way ANOVA, n = 3). (B1) 2.5e4 Pzp53Med cells irradiated at 2.5Gy and 5Gy, harvested after 3 days, and counted (3.45e6 shSCR NT vs 2.12e6 shYB1 NT p < 0.0001; 2.42e6 shSCR 2.5Gy vs 1.31e6 shYB1 2.5Gy p < 0.0001; 1.11e5 shScr 5Gy vs 5.1e4 shYB1 5Gy p = 0.0039, n = 3). (B2) Doubling time calculated for Pzp53Med (10.34 h shSCR NT vs 11.04 h shYB1 NT p = 0.2149; 10.85 h shSCR 2.5Gy vs 12.37 h p = 0.0036; 12.84 h shSCR 5Gy vs 16.95 h shYB1 5Gy p < 0.0001, n = 3). (C) β-Gal stain of UW228 cells following radiation time course demonstrating increased senescence of YB1 depleted cells compared to irradiated control (5Gy shSCR vs 5Gy shYB1 95% CI = −16.75–(−5.09) p < 0.0001 two-way ANOVA, n = 3). (D1) 2.5e5 D341 cells plated and irradiated followed by a 5 day incubation period (2.6e6 shLUC NT vs 1.48e6 shYB1 NT p = 0.0114; 8.0e5 shLUC 2.5Gy vs 6.13e5 shYB1 2.5Gy p = 0.0450; 3.3e5 shLUC 5Gy vs 2.1e5 shYB1 5Gy p = 0.0407, n = 3). (D2) Doubling time for D341 (35.50 h shLUC NT vs 46.78 h shYB1 NT p = 0.0074; 71.83 h shLUC 2.5Gy vs 92.76 h p = 0.0001, n = 3). (E1) 2.5e5 D425 cells plated and irradiated followed by a 5 day incubation period (1.58E6 shSCR NT vs 6.25E5 shYB1 NT p < 0.0001; 8.97e5 shSCR 2.5Gy vs 3.02e5 shYB1 2.5Gy p < 0.0001; 4.43e5 shSCR 5Gy vs 1.85e5 shYB1 5Gy p = 0.0124, n = 3). (E2) Doubling time for D425 (45.2 h shSCR NT vs 93.7 h shYB1 NT p = 0.0281; 68.1 h shSCR 2.5Gy vs 575.4 h p = 0.0140, n = 3). All comparisons performed using 2-way ANOVA, see Supp Fig. 11 for growth and doubling times statistics.
