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. 2021 Jul 27;7:196. doi: 10.1038/s41420-021-00574-1

Fig. 4. TRIB2 reduced labile iron level in a TFRC-dependent manner.

Fig. 4

A Schematic presentation of iron metabolism. B mRNA levels of indicated iron regulators before and after knocking down by siRNA, as measured by qPCR. C Deletion of TRIB2 elevated labile iron via TFRC in Bel-7404 cells. Labile iron was measured by Iron Assay Kit before and after TRIB2 was knocked out in Bel-7404 cells with siRNA-mediated knockdown for indicated iron regulators. Deletion of TRIB2 elevated labile iron via TFRC. TFRC and TRIB2 were measured by IB (D). Labile iron was measured by Iron Assay Kit (E) in Bel-7404 cells with or without single or double knockout (DKO) of TRIB2 and TFRC in basal condition or following treating with RSL3 (5 µM, 12 h) or erastin (10 µM, 12 h). F, G Overexpression of TRIB2 reduced labile iron also via TFRC. TFRC and TRIB2 were measured by IB (F). Labile iron was measured by Iron Assay Kit (G) before and after overexpressing TRIB2 in SK-Hep1 cells with or without TFRC knocked out in basal condition or following treating with RSL3 (5 µM, 12 h) or erastin (10 µM, 12 h). Data were analyzed by Student’s t test and expressed as mean ± SD from three independent experiments. NS non-significance; **P < 0.01; ***P < 0.001; ****P < 0.0001. Images of all the immunoblots are representative of three independent experiments.