Fig. 2: Naphthofluorescein disrupts Mint3–FIH-1 interaction and attenuates HIF-1 activity.
From: Pharmacological inhibition of Mint3 attenuates tumour growth, metastasis, and endotoxic shock
a Pull-down assay of GST-FIH-1 and His6-Mint3 in the presence of DMSO or naphthofluorescein (Naph; 10 μM). b Mint3 expression in HT1080 cells transfected with siRNAs against green fluorescent protein (GFP; siGFP) and Mint3 (siMint3#1, #2). c Luciferase assay for HIF-1 activity in siRNA-transfected HT1080 cells treated with DMSO or Naph (10 μM). d V5-tagged Mint3 expression in HT1080 cells transfected with mock or V5-tagged Mint3 expression vector. e Luciferase assay for HIF-1 activity in mock or V5-tagged-Mint3 expressing HT1080 cells treated with DMSO or Naph (10 μM). f Mint3 expression in HT1080 cells transfected with siRNAs against GFP (siGFP) and FIH-1 (siFIH-1#1, #2). g Luciferase assay for HIF-1 activity in siRNA-transfected HT1080 cells treated with DMSO or Naph (10 μM). h–n mRNA levels of glycolysis-related HIF-1 target genes (h–l) and HIF1A (m), and 18S rRNA levels (n) in HT1080 cells treated with DMSO or Naph at the indicated concentration for 24 h. o Cell growth assay of HT1080 cells treated with DMSO or Naph at the indicated concentrations. In (c), (e), (g–o), error bars indicate SD (n = 3). Data were analysed using Student’s t test. *p < 0.05, **p < 0.01, ***p < 0.001. NS not significant.
