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. 2023 Jul 27;14(7):475. doi: 10.1038/s41419-023-06008-3

Fig. 5. THOC3 promotes LUSC partially through PFKFB4 by binding to YBX1.

Fig. 5

A Cell lysates from H1703 cells transfected with Flag-tagged THOC3 were immunoprecipitated with anti-YBX1, anti-Flag, or IgG antibodies, and the immunoprecipitates were blotted with anti-Flag/YBX1 antibodies. B Immunofluorescence shows the distribution of THOC3 (green) and YBX1 (red) in H1703. Scale bars = 100 μm. C Genes significantly downregulated upon THOC3 or YBX1 knockdown (KD) and associated with the carbohydrate metabolism (Venn diagram). D Outcomes of qRT-PCR and WB showing PFKFB4 expression in LUSC cells transfected with si-NC or si-THOC3/YBX1. E PFKFB4 levels in B2B, SK-MES-1, H1703, and H520 are shown using qRT-PCR and WB (left); PFKFB4 expression in cells transfected with scramble or THOC3-OE plasmids shown by WB (right). F, G Growth curves (day 1–4) and colony formation assays show the proliferation of H520/H1703 cells transfected with si-NC or PFKFB4-si1/2. H Transwell assays were performed to assess the migration of H520/H1703 cells with PFKFB4 knockdown. Cells crossing the membrane were dyed with crystal violet (10× magnification). Scale bars = 100 μm. I The relative glucose uptake, lactic acid and ATP production of H520/H1703 cells transfected with si-NC or PFKFB4-si1/2 after 24 h. *P < 0.05; **P < 0.01; ***P < 0.001. Variables are presented as mean ± SD.