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. 2018 Apr 4;13(4):e0195050. doi: 10.1371/journal.pone.0195050

Fig 3. V158411 does not thermostabilize Chk1 in intact cells heated by the PBS immersion method.

Fig 3

(A) Mean nuclear Chk1 intensity was determined in U2OS and HT29 cells following immunofluorescent staining with the anti-Chk1 antibodies G-4 and EP691Y and high content imaging. Mean of 3 independent wells ± SD. (B) HT29 cells were transiently transfected with control (siCtrl) or CHEK1 smart pool siRNA (siCHEK1) for 48 hours. Mean nuclear Chk1 intensity was determined by immunofluorescent staining with G-4 or EP691Y antibodies and high content imaging. (C) Thermal response profile of Chk1 in HT29 cells detected by immunofluorescent imaging with the EP691Y antibody following immersion in PBS pre-heated to the indicated temperature. Values are the mean of four wells ± SD. (D) HT29 cells were treated with 0–10 μM V158411 for 30 minutes before being heated by immersion in PBS pre-heated to the indicated temperature for 3 minutes followed by room temperature for 5 minutes. Nuclear Chk1 intensity was determined by immunofluorescent staining using the EP691Y antibody and high content imaging. Red line, mean.