Fig 1: Low SLC7A2 upregulates CXCL1 expression by transactivating P65 expression.A Western Blotting the SLC7A2, phosphorylated P65 at ser536 (p-P65Ser536), P65, P50, p-IKBa, IKBa, ß-actin was used as control. B qRT-PCR was used to detect that relative mRNA expression of CXCL1 in the indicated cells with treatment with NF-k?B inhibitor, JSH-23 (10 µm, 24 h). The data are shown as the mean ± SD from the at least three independent experiments; *P < 0.05, **P < 0.01. C ELISA was used to detect that relative expression of CXCL1 in the indicated cells with treatment with NF-?B inhibitor, JSH-23 (10 µm, 24 h). The data are shown as the mean ± SD from the at least three independent experiments; *P < 0.05, **P < 0.01. D The expression of phosphorylated P65 was analyzed in the nuclear and cytoplasmic fractions of Huh7-shSLC7A2 cells and MHCC97H-SLC7A2 cells. The laminB1 and ß-actin were as control respectively. E Luciferase reporter assay was showed in the indicated co-transfected with pCMV-SLC7A2 and the CXCL1 promoter-luciferase construct. The data are shown as the mean ± SD from the at least three independent experiments; **P < 0.01. F Truncated and mutated CXCL1 promoter constructs were co-transfected with PCMV-SLC7A2, and the relative luciferase activity was confirmed. The data are shown as the mean ± SD from the at least three independent experiments; *P < 0.05. G ChIP assay confirmed the direct binding of P65 to the CXCL1 promoter in HCC cells. The data are shown as the mean ± SD from the at least three independent experiments; *P < 0.05. H ChIP assay confirmed the direct binding of P65 to the CXCL1 promoter in human HCC tissues. The data are shown as the mean ± SD from the at least three independent experiments; *P < 0.05. I Immunofluorescence of p-P65Ser536 of indicated HCC cells. J Huh7-shSLC7A2 cells treated with inhibitors of PI3K, JNK, ERK, p38, STAT3, and then western blotting was used to detect the expression of SLC7A2 and p-P65Ser536 in the nuclear as well as the total and phosphorylate expression of AKT, ERK, JNK, p38, STAT3.