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. 2020 Oct 12;21(20):7523. doi: 10.3390/ijms21207523

Figure 3.

Figure 3

Effect of EGR-1 silencing on CYP19 expression. (A) EGR-1 expression in stably transfected MCF-7 cells expressing scrambled control (shCT) or EGR1 short-hairpin RNA (shEgr1) was verified by immunoblotting. Exponentially growing MCF-7 variant cells were cultured in the presence or absence of 10 ng/mL TNFα for 1 h. The cells were then harvested and analyzed by immunoblotting using anti-EGR-1 antibody. GAPDH was used as an internal control. (B–D) MCF-7 cells expressing shCT or shEgr1 were treated with 10 ng/mL TNFα for 24 h. CYP19 mRNA levels were determined by RT-PCR (B) and qR-PCR (C). GAPDH mRNA levels were used as an internal control. CYP19 aromatase levels were measured by immunoblot analysis (D). GAPDH levels were used as an internal control. Band intensities were measured using the ImageJ software. Bars represent the mean ± SD (n = 3). * p < 0.05, *** p < 0.001 by Dunnett’s multiple comparisons test. NS, not significant; qR-PCR, quantitative real-time PCR.