Fig. 6

Transcriptional regulation of SLC2A1 by GDF15-TGFβ signaling cascades in MM. (A) In H929 cells, real-time qPCR analysis showed the effects of GDF15 overexpression or rGDF15 on the mRNA expression of SLC2A1 in the presence or absence of LY2109761 (n = 3 per group). (B) In H929 cells, western blotting showed the effects of GDF15 overexpression or rGDF15 on the protein expression of SLC2A1 in the presence or absence of LY2109761. (C) CHIP experiment analysis showed the enrichment of Smad2 in the different promoter region of SLC2A1 gene. (D) Luciferase reporter assay showed the promoter activity of different segments upon rGDF15 stimulation (n = 3 per group). (E-F) Real-time qPCR and western blotting analysis showed the knockdown efficiency of SMAD2 gene and its effects on the mRNA expression of SLC2A1 (n = 3 per group). (G) Luciferase reporter assay showed the SLC2A1 promoter activity upon GDF15 overexpression and SMAD2 knockdown (n = 3 per group). All experiments were repeated two times. P values were calculated using unpaired two-tailed Student’s t-test or one-way ANOVA for Tukey’s multiple-comparisons test. *P < 0.05, **P < 0.01, ***P < 0.001