Fig. 4

SH3GLB1 downregulation reduces TMZ-enhanced autophagy. A The SH3GLB1 protein (Red) is colocalized (white arrow) with autophagosomes labeled by LC3-EGFP (Green). The statistic graph showing the numbers of autophagic puncta (dot). Scale bar = 50 μm (B) Resistant cells treated with shSH3GLB1 exhibits attenuated autophagy levels. C The parental cells with SH3GLB1 overexpression exhibit enhanced autophagy levels. D Western blot analysis showing the expression of LC3, p62, Atg5–12 complex, and OXPHOS complexes from cells with or without SH3GLB1 knockdown that were cotreated with chloroquine (CQ). E Western blot analysis showing increased levels of cleaved caspase 3 in recurrent GBM spheroid cultures with siSH3GLB1 and TMZ cotreatment for 24 hours. F Autophagy enhanced by evidence of altered LC3B-II and p62 expression after TMZ treatment for 24 hours was attenuated in siSH3GLB1 cells. G Attenuated detection of induced acridine orange–stained acidic vesicular organelles is shown following TMZ treatment (24 hours) and siSH3GLB1 knockdown. The statistical graph obtained by flow cytometry is shown in the right panel (Scale bar = 200 μm; *p < 0.05; N = 3 in each group; R: resistance)