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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: Hypoxia induced ferritin light chain (FTL) promoted epithelia mesenchymal transition and chemoresistance of glioma

Fig. 2

Hypoxia induced FTL in a HIF-1α dependent manner. a Representative images of IHC staining of FTL and HIF1A in glioma tissues and Chi-square test was used for comparison between groups b, Scale bars,20 μm; c Ivy glioblastoma atlas project (Ivy GAP) was used to analyzed FTL expression in different anatomic areas. LE, leading edge; IT, Infiltrating tumor; CT, cellular tumor; Pan, pseudopalisading cells around necrosis; Mvp, microvascular proliferation; d U87 and U251 cells were cultured under hypoxia and FTL expression was assessed by western blot. HIF1A was used as positive control.β-actin was used as loading control; e U251 and U87 cells were cultured under hypoxia. Whole cell lysates were collected and western blot was performed to find dynamic change of HIF1A,HIF2A and FTL protein expression.f Glioma cells were treated with CoCl2(400 mM) for 24 h,and HIF1A HIF2A, FTL expression were determined by Western blot.g-i Cells were transfected with siRNA-HIF1A or siRNA-HIF2A.mRNA and protein level of FTL,HIF1A and HIF2A were measured by RT-PCR and western blot, respectively. j Show for ChIP-seq profiles of HIF1A and HIF1A for a region of chromosome 19 obtained using RNAseq from Hela and T47D cells. Position of POLRA2A was used as positive control. k Putative hypoxia response elements (HREs) on the region of FTL promoter and graphic representation of mutant FTL promoter. L FTL promoter activity of U87 and U251 cells detected by Luciferase reporter assay in hypoxic or normoxic condition. Besides, m mutant-FTL-Luc or wildtype-FTL-Luc plasmids were then co-transfected with the Renilla luciferase reporter plasmid into glioma cells. Hypoxia maintenance time is 24 h. n and o Representative image of ChIP of HIF1A binding to FTL promoter under hypoxia. Anti-HIF1A and anti-IgG antibodies were used. All image represented as the mean ± SD of three independent experiments; *,P < 0.5;**,P < 0.01;***,P < 0.001, ns, no significance

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