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. 2013 Sep 27;8(9):e75568. doi: 10.1371/journal.pone.0075568

Figure 3. GDF15 is responsible for K7174-mediated HAMP gene downregulation in HepG2 cells.

Figure 3

(A) Retrovirus-mediated GDF15 overexpression in HepG2 cells. Anti-GDF15 antibody was used to detect endogenous GDF15 protein. Alpha-Tubulin was used as a loading control. (B) Quantitative RT-PCR analysis of HAMP expression in GDF15 overexpressed HepG2 cells. The expression level of each target gene relative to that of GAPDH was calculated (n = 3, mean ± SE, * P < 0.05). The expression level of DMSO-treated control cells was set to 1. (C) Experimental strategy for siRNA-mediated GDF15 knockdown after K7174 treatment in HepG2 cells. (D) Western blotting analysis of whole-cell extracts from GDF15-silenced HepG2 cells, treated with K7174 or DMSO. Anti-GDF15 antibody was used to detect endogenous GDF15 protein. Alpha-Tubulin was used as a loading control. (E) Quantitative RT-PCR analysis of GDF15 and HAMP expression in GDF15-silenced HepG2 cells, treated with K7174 or DMSO. The expression level of each target gene relative to that of GAPDH was calculated (n = 3, mean ± SE, * P < 0.05). The expression levels of DMSO- and control siRNA-treated control cells were set to 1.