Fig. 5
ROCK2’s crosstalk with DNA repair machinery in cervical cancer cells. a Immunoblot of irradiated SiHa cells showing expression levels of DNA repair proteins at various time points indicated, as compared to control at 0 h. b-i Representative immunofluorescence images depict the co-expression of ROCK2 and pH2Ax in CaSki cells at 1 h following irradiation (n = 3, scale bar = 10 μm). b-ii Representative immunofluorescent images of irradiated CaSki cells showing a decreased expression of pH2Ax in ROCK2In as compared to IgGIn cells (n = 3, scale bar = 10 μm). b-iii Box plots of the number of pH2Ax foci in IgGIn and ROCK2In cells following irradiation. A median value of 15 foci/cell in IgGIn as opposed to 5 foci/cell in the ROCK2In was observed (n = 3, *p < 0.01). b-iv Immunoblot analysis demonstrated a reduction in pH2Ax levels at 1 h in irradiated ROCK2In as compared to IgGIn cells (n = 3). b-v Immunoprecipitation using ROCK2 antibody shows pull down of pH2Ax at 1 h following irradiation treatment. IgG isotype is used as the control (n = 3). c-i Representative immunofluorescence images showing a decreased expression of RAD50 as compared to IgGIn, in irradiated CaSki cells, with ROCK2In (n = 3, scale bar = 10 μm). c-ii Immunoblot analysis in irradiated SiHa cells confirming reduction in RAD50 levels upon ROCK2In (n = 3). d-i Representative immunofluorescence images showing a decreased expression of MRE11 as compared to IgGIn cells, in irradiated CaSki cells, with ROCK2In (n = 3, scale bar = 10 μm). d-ii Immunoblot analysis of irradiated SiHa cells also showed a reduction in MRE11 levels upon ROCK2In (n = 3)