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. 2018 Jun 29;19(7):1909. doi: 10.3390/ijms19071909

Figure 4.

Figure 4

(A) CH-5 affects the expression of Sp1, p53, and DNMT1 proteins in U2OS cells. The cells were grown in a 60 mm dish and then were incubated with CH-5 at the indicated concentrations for 24 h. A 30 µg aliquot of total proteins was examined by western blotting, as described in Materials and Methods; (B) Effect of CH-5 and curcumin on the expression of DNMT1 mRNA, assessed by RT-PCR. U2OS cells were treated with DMSO, CH-5 10, 20, and 40 µM, or curcumin 30 µM for 24 h. The transcript levels were normalized using RPL30 as a reference gene; * p < 0.05 and ** p < 0.01; (C) Effects of CH-5 treatment and Sp1 overexpression on the mRNA levels of DNMT1. When U2OS cells were simultaneously exposed to CH-5 (40 µM) and transfected with an Sp1-expressing vector, there was a weaker downregulation of DNMT1, compared to control cells transfected with a control empty vector. The data are indicated as fold change in relative expression compared with RPL30 as a reference gene and on the basis of the comparative ΔΔCt method, and expressed as the mean ± standard deviation; * p < 0.05 and ** p < 0.01.