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Fig. 4 | Cell & Bioscience

Fig. 4

From: A physical association between the human mutY homolog (hMYH) and DNA topoisomerase II-binding protein 1 (hTopBP1) regulates Chk1-induced cell cycle arrest in HEK293 cells

Fig. 4

Knockdown of both hTopBP1 and hMYH affects cell cycle arrest. a HEK293 cells were either transfected with individual siRNAs targeted against GFP (siGFP), hMYH (siMYH), and hTopBP1 (siTopBP1) or co-transfected with siTopBP1 and siMYH. Transfected cells were treated with 20 mM HU for 1 h. Total cell lysates were immunoblotted with anti-hTopBP1, anti-hMYH, anti-Chk1, and anti-phospho-Chk1 antibodies. b The expression of hTopBP1, hMYH, or both was knocked down, following which the cells were incubated with 20 mM HU for 1 h. The cell lysates were analyzed by immunoblotting with anti-p-Cdk2 (T24, Y15) and anti-Cdc25A antibodies. c The relative levels of p-Chk1, Cdc25A, and p-Cdk2 from replicate experiments were quantified and normalized to β-actin, and the normalized data are presented as mean ± standard error (of three independent experiments). P values were calculated using a paired t test, *p < 0.05, **p < 0.01, ***p < 0.001

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