As shown in Figure 1A, Hela and Siha cells transfected with DNMT1-siRNA (transfection group) displayed lower level of mRNA expression (P < 0.01), with inhibitory ratios of 56.21% and 41.31% respectively compared with control group (negative siRNA). No significant change in DNMT1 mRNA expression was found between control group and blank control
(Lipo 2000). The transcript https://www.selleckchem.com/products/lcz696.html quantity of GAPDH in transfection group, control group and blank control did not change significantly. Figure MAPK inhibitor 1B showed the DNMT1 protein expression levels in Hela and Siha cells at 72 h after transfected with DNMT1-siRNA. The protein level of DNMT1 decreased significantly compared with control group and blank control (P < 0.01). The inhibitory ratios of DNMT1 protein level in Hela and Siha cells were 50.31% and
99.76%, respectively. Figure 1 Effects of siRNA on DNMT1 mRNA and protein expression. (A): mRNA expression levels of DNMT1 in Hela and Siha cells were examined by qPCR. Compared with control group, Hela and Siha cells Epacadostat cell line transfected with DNMT1-siRNA displayed lower level of mRNA expression (**P < 0.01). (B): DNMT1 protein levels in Hela and Siha cells were determined by western blot. The protein level of DNMT1 decreased significantly compared with control group and blank control. (1: transfection group (DNMT1-siRNA); 2: control group (negative siRNA); 3: blank group (Lipo2000), n = 3). Effects of DNMT1 silencing on cell cycle and apoptosis The G0/G1 ratio (74.72 ± 3.17%) of Hela cells in transfection group was higher than that in control group (65.88 Liothyronine Sodium ± 3.23%) (P < 0.01), and cells at S phase were fewer compared with control group. Meanwhile, The G0/G1 ratio (76.43 ± 2.20%) of Siha cells in transfection group displayed significantly higher compared with control group (66.4 ± 1.99%) (P < 0.01), while cells at S phase were fewer than those in control group. No significant changes in G0/G1 ratio or cells at S phase were detected between the control group and blank control (Figure 2A). Furthermore, as shown in Figure 2B, the apoptosis of Hela cells in transfection group was significantly higher than that
in control group (P < 0.01). Similar results were observed in Siha cells. Figure 2 Effects of DNMT1 silencing on cell cycle and apoptosis. (A): Phases of cell cycle of Hela and Siha cells were analyzed by flow cytometry assay at 48 h after transfection (**P < 0.01). (B): Apoptosis of Hela and Siha cells was analyzed by flow cytometry assay at 48 h after transfection (**P < 0.01). (1: transfection group (DNMT1-siRNA); 2: control group (negative siRNA); 3: blank group (Lipo2000), n = 3). Effects of DNMT1 silencing on cell growth and proliferation Cell growth and proliferation of Hela and Siha cells were examined using MTT assay. As shown in Figure 3, viabilities of Hela cells in transfection group were 91.47%, 86.74%, 78.