Figure 2

Depletion of UHRF1 or DNMT1 enhances sensitivity to PMA and upregulates cytokines related to M1 polarization in THP-1 cells. (a) FACS analysis to determine the counts of CD14-positive cells. Control and UHRF1 knockdown cells were treated with DMSO or PMA for 48 h. Data are shown as mean ± SEM (n = 3). The P-values were calculated by one-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01 and ****P < 0.0001. (b) FACS analysis to determine the counts of CD14-positive cells. Control and DNMT1 knockdown cells were treated with DMSO or PMA for 48 h. Data are shown as mean ± SEM (n = 3). The P-values were calculated by one-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01 and ****P < 0.0001. (c) FACS analysis to determine the counts of CD14-positive cells. Control and DNMT3B knockdown cells were treated with DMSO or PMA for 48 h. Data are shown as mean ± SEM (n = 3). The P-values were calculated by one-way ANOVA followed by Tukey’s multiple comparisons test. **P < 0.01, ***P < 0.001 and n.s., not significant. (d) qRT-PCR analysis to determine the mRNA level of TNF, IL1B and IL6 in PMA-treated THP-1 cells. Cells were treated with PMA for 48 h and incubated for additional 48 h before harvest. Data are shown as mean ± SEM (n = 3). The P-values were calculated by one-way ANOVA followed by Dunnetts’s multiple comparisons test. *P < 0.05, **P < 0.01 and n.s., not significant. (e) Relative abundance of secreted TNF-α, IL-1β, IL-6 were analyzed by ELISA. Cells were treated with PMA for 48 h and incubated for additional 48 h before harvest. Relative absorbance at 450 nm were used for plotting. Data are shown as mean ± SEM (n = 3). The P-values were calculated by one-way ANOVA followed by Dunnetts’s multiple comparisons test. **P < 0.01, ****P < 0.0001 and n.s., not significant.