Fig. 3: SETD8 inhibition by siRNAs induces DNA damage and cell cycle arrest in glioblastoma cell lines. | Cell Death & Disease

Fig. 3: SETD8 inhibition by siRNAs induces DNA damage and cell cycle arrest in glioblastoma cell lines.

From: Targeted inhibition of the methyltransferase SETD8 synergizes with the Wee1 inhibitor adavosertib in restraining glioblastoma growth

Fig. 3

a LN-18 and U251 cells were transfected with non-targeting siRNAs (siCTRL), with an siRNA pool targeting the SETD8 3′-UTR (siSETD8), alone or with also a plasmid expressing an siRNA resistant SETD8, pcDNA myc-Flag SETD8 (siSETD8+rescue). Cells were fixed and immunostained for p-γ-H2AX (red) to assess DNA damage and for β-tubulin (green) to label the cytoskeleton and ensure cellular integrity. DNA was stained by DAPI (blue). Representative fluorescent microscopy images are shown (scale bars: 20 μm). Graphs show percentage of cells positive for p-γ-H2AX foci (cells were called positive if with ≥5 foci). Statistical analyses were performed using Student’s t test. b A fraction of siCTRL, siSETD8 and siSETD8+rescue cells were collected and lysed. Extracted proteins were separated on SDS-PAGE, blotted and probed for the indicated antigens. All values are given as mean ± standard deviation of at least three replicates. *** p < 0.001; ** p < 0.01; * p < 0.05.

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