Skip to main content
. 2021 Feb 9;11(2):246. doi: 10.3390/biom11020246

Figure 4.

Figure 4

Evaluation of intracellular EPI accumulation in MDA-MB-231 cells after knockdown of UGDH gene. MDA-MB-231 cells were transfected with a specific siRNA against UGDH gene (siUGDH) using a random sequence siRNA as the negative control (siSCR). After 24 h, 1 µM EPI (EPI) was added to complete 48 h of incubation. Intracellular EPI accumulation was measured by flow cytometry. A control with MDA-MB-231 cells treated only with EPI (EPI) was added to determine the basal uptake of the drug. Bars show the percentage of EPI+ cells determined by comparison with a negative control (A). Histograms (B) and dot plots (C) show the most representative of four independent experiments performed with 50,000 events/condition. n = 4, mean ± SEM, * p < 0.05. To evaluate the expressions of ABCG2 (D), ABCC1 (E), ABCC2 (F), UGT2B7 (G), drug efflux pumps and the specific EPI transferase UGT2B7, total RNA was extracted and 2 µg was reverse transcribed by RT-PCR. cDNAs were subjected to real-time quantitative PCR using SYBR Green. Results were normalized using β-actin as reference gene and all determinations were performed as duplicates. n = 3, mean ± SEM, * p < 0.05 ** p < 0.01 *** p < 0.001.