Fig. 2
TFRC knockdown inhibited cell growth in vitro. (A) Successful knockdown of TFRC levels using two different siRNAs in HK1-EBV cells. (B) The intracellular Fe2+ visualized using FerroOrange was decreased after TFRC knockdown compared to the level in control cells. (C) Knockdown of TFRC inhibited the iron storage protein FTH1. (D) The CCK-8 assays showed knockdown of TFRC significantly suppressed cell proliferation from day 2 compared to that in control cells. (E) Representative images from colony formation cells, and the quantitative data indicate that knockdown of TFRC inhibited the colony formation ability in NPC cell line. (F) Representative distribution of the apoptotic cells using the Muse cell analyzer, and the quantitative data indicating total apoptotic cells significantly increased in siTFRC#1(23.9%) and siTFRC#2(22.7%) than that in the control (17.1%). (G) Knockdown of TFRC inhibited anti-apoptosis protein BCL-xL. (H) Representative cell cycle plots for the cells using Muse™ Cell analyzer, and the quantitative data indicating a significant arrest of siTFRC#1 group (54.4%) and siTFRC#2(58.4%) cells in the G0/G1 cell cycle phases compared to that in the control (43.5%). The results are given for three independent experiments. Asterisks indicate *p < 0.05, **p < 0.01, ***p < 0.001 compared to control