Fig. 4: HABON interacted with VDAC1 on mitochondria.
From: LncRNA HABON promoted liver cancer cells survival under hypoxia by inhibiting mPTP opening
A 4XS1m-HABON or 4XS1m was expressed in Huh7 cells under hypoxia treatment for 24 h. Use streptavidin magnetic beads for immunoprecipitation experiments. Mass spectrometry were performed after protein gel electrophoresis and the binding protein were further verified by western blot. B Use the IPA (Ingenuity Pathway Analysis) bioinformatics website to perform functional enrichment analysis on the protein binding with HABON. C Huh7 cells were treated with normoxia and hypoxia for 24 h. After the samples were fixed, HABON RNA FISH and VDAC1 immunofluorescence experiments were performed, and they were observed under a confocal microscope. The scale bar is 18.4 μm. D Isolation of mitochondria in Huh7 cells cultured under normoxia and hypoxia for 24 h. qRT-PCR was used to detect the expression of HABON in the mitochondrial components. E 4XS1m-HABON or 4XS1m was expressed in Huh7 cells with hypoxia treatment for 24 h. Use streptavidin magnetic beads for immunoprecipitation experiments. Western blot verified the combination of HABON with VDAC1. F Huh7 cells were cultured under hypoxia for 24 h an RIP experiment was performed with anti-VDAC1 antibody or IgG. qRT-PCR detected the enrichment of VDAC1 on HABON and negative controls Actin, GAPDH and U6 in RNA samples. G Schematic of the colocalization of HABON with VDAC1 on outer mitochondrial membrane. Error bars stand for mean ± SD. Three independent experiments, two-tailed Student’s t-test. p < 0.05, **p < 0.01, ***p < 0.001.
