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. 2022 Mar 2;13(3):199. doi: 10.1038/s41419-022-04647-6

Fig. 2. ALDOA interacts with HIF-1α and STAT3-ALDOA mediates ATP-HIF-1α signaling.

Fig. 2

A MCF-7 cells lysed and co-immunoprecipitated (co-IP) with anti-immunoglobulin G (IgG) or anti-HIF-1α. elutes were resolved by SDS-PAGE and silver-stained (left). The presence of ALDOA was detected by mass spectrometry and demonstrated by western blotting (right). B Physical interaction between ALDOA and HIF-1α was determined by co-immunoprecipitation and western blotting in both MDA-MB-231 and MCF-7 cells. C The interaction between ALDOA and HIF-1α was demonstrated by GST pull-down assays with bacterially expressed GST-fused ALDOA and endogenous transcribed/translated HIF-1α (left), or with GST-fused HIF-1α and endogenous transcribed/translated ALDOA (right). D MDA-MB-231 and MCF-7 cells were double-stained with ALDOA (green) and HIF-1α (red), counterstained with DAPI (blue), and observed under a confocal microscope. The merged regions indicated their co-localization. E Western blotting proved that the elevation of HIF-1α via ATP was attenuated by ALDOA-siRNAs in MDA-MB-231 and MCF-7 cells. F Western blotting illustrated that the expression changes of ATP-HIF-1α signaling were attenuated by S3I-201 (STAT3 inhibitor) in MDA-MB-231 and MCF-7 cells. Data are representative of at least three independent experiments. Error bars represent means ± SD from triplicate experiments. *p < 0.05; **p < 0.01; ***p < 0.001; ns, not significant.