Fig. 4

Aggressive phenotype in cases exhibiting MASI of H3F3A K27M. a Immunofluorescence staining of H3 K27M (red) and histone H4 (green) as an internal control of MASI case (case 5; bars, 30 μm) and lower VAF case (case 6). b Bar graph of relative fluorescence intensity of H3 K27M to those of histone H4 in lower VAF and MASI cases (n = 6 and n = 4, respectively). Error bars indicate the standard deviation (SD) (**P < 0.05). (c) Bar graph indicates the frequency of H3K27me3 positive cells among histone H4 positive cells in lower VAF and MASI cases (n = 6 and n = 4, respectively). Error bars indicate the SD (***P < 0.01). d Immunofluorescence staining of Ki-67 (green), H3 K27M (red), and DAPI (blue) using the tumor specimen of the MASI case (case 5; bars, 20 μm) and lower VAF case (case 4). e Bar graphs indicating the frequency of Ki-67 positive cells among H3 K27M positive cells in lower VAF cases (n = 6, blue) and MASI cases (n = 4, red). Error bars indicate the SD (**P < 0.05). f Kaplan-Meier survival curves illustrating the progression-free survival (PFS) of MASI cases (n = 4) and lower VAF cases (n = 11; P = 0.03). g Kaplan-Meier survival curves illustrating the overall survival (OS) of MASI cases (n = 4) and lower VAF cases (n = 11; P = 0.01)