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. 2018 May 10;13(5):e0197165. doi: 10.1371/journal.pone.0197165

Fig 6. The knock-down of DBP5, GLE1 or IPPK alters DNA damage response.

Fig 6

(A) U2OS cells transfected with siRNA of DBP5, GLE1 or IPPK were cultured for 48 h. After fixation, immunostaining was performed using anti-γH2A.X antibody. Chromosome was counterstained with DAPI. Scale bar, 20 μm. (B) The nuclear signal intensity of γH2A.X was quantified in each knock-down cell. Each value is the mean with SD of three independent experiments. Error bars represent the SD. p-values were calculated by one-way ANOVA followed by Dunnett’s test by comparison with the control. (n = 30, *** = p<0.001). (C) Immunoblotting against H2A.X was performed using the nuclear extract fraction of U2OS cells transfected with indicated siRNA. Lamin b was used as a loading control. (D) Real-time PCR was performed to detect cytoplasmic mRNA amounts of factors belonging to the DNA damage response category in IPA. Cytoplasmic total RNA was recovered from U2OS cells transfected with indicated siRNA. White bar: the detected value of real-time PCR. Gray bar: the detected value of microarray analysis. Each value is the mean with SD of three independent experiments. Error bars represent the SD. p-values were calculated by one-way ANOVA followed by Dunnett’s test by comparison with the control. (*** = p<0.001, ** = p<0.01 and * = p<0.05).