Fig 1: CLIC1 expression in HCC cells affected VEGFA secretion. (A) Validation of stable CLIC1-knockdown and -overexpressing in Huh7 cell lines. (B) Validation of stable CLIC1-knockdown and -overexpressing in BEL 7402 cell lines. (C) The concentrations of VEGFA in Huh7 cell lines. (D) The concentrations of VEGFA in BEL 7402 cell lines.
Fig 2: Overexpression of mTOR can rescue ESCC cells from CLIC1 mediated inhibition of cell proliferation. A-B. KYSE150 and ECA109 cells were transfected with siCtrl or si-CLIC1 and then transfected with vector pcDNA3.1 or plasmid for m-TOR overexpression. The indicated proteins were detected by Western blotting, and ß-actin was used as loading control. C-D. CCK-8 assays of cell proliferation in A and B. Mean ± SEM. n=3. *P < 0.05.
Fig 3: Chloride efflux is essential for NLRP3 inflammasome activation. a–c ELISA of IL-1ß in LPS-primed BMDMs which were transferred to a basic NaCl saline (130 mM NaCl, 5 mM KCl, 1 mM MgCl2, 1 mM CaCl2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose) or chloride-free NaGluc saline (130 mM NaGluc, 5 mM KGluc, 1 mM MgGluc2, 1 mM CaGluc2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose) and left stimulated with ATP a, MSU b, or nigericin c. d ELISA of IL-1ß in LPS-primed BMDMs which were transferred to NaCl saline, NaBr saline (130 mM NaBr, 5 mM KBr, 1 mM MgBr2, 1 mM CaBr2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose), NaI saline (130 mM NaI, 5 mM KI, 1 mM MgI2, 1 mM CaI2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose), NaGluc saline or NaGlut (130 mM NaGlut, 5 mM KGlut, 1 mM MgGlut2, 1 mM CaGlut2, 20 mM HEPES (pH 7.4), 1 mg/ml BSA, 10 mM glucose) saline for different time points. e, f Immunoblot analysis IL-1ß and cleaved caspase-1 e or ELISA of IL-1ß in supernatants f of LPS-primed Nlrp3 +/+ or Nlrp3 –/– BMDMs which transferred to NaCl, NaBr or NaI saline for 3 h. g ELISA of IL-1ß in supernatants of LPS-primed Clic4 –/– BMDMs transfected siRNA against Clic1 and Clic5 and left stimulated with nigericin for 30 min or transferred to NaCl, NaBr or NaI saline for 3 h. Data are from three independent experiments with biological duplicates in each (a–d, f, g; mean ± SEM of n = 6) or are representative of three independent experiments e. Student’s t-test a–c, f, g, ***P < 0.001
Fig 4: Clics-dependent chloride efflux during NLRP3 inflammasome activation. a–c Qualification of the decrease of intracellular chloride in wild type or Nlrp3 –/– BMDMs at different time points after nigericin a, ATP b, or poly A:T c treatment. d, e Qualification of the decrease of intracellular chloride in BMDMs at different time points after nigericin d or ATP e treatment with or without the pretreatment with IAA94 (150 µM). f, g Qualification of the decrease of intracellular chloride in Clic4 –/– BMDMs transfected siRNA against Clic1 and Clic5 at different time points after nigericin f or ATP g treatment. Data are from three independent experiments with biological duplicates in each (mean ± SEM of n = 6). Student’s t-test, *P < 0.05, **P < 0.01, ***P < 0.001
Fig 5: CLIC1 expression in HCC cells affected the migration ability of endothelial cells. (A) EA.hy926 cells were coculture with HCC cells with different expression levels of CLIC1. (B) EA.hy926 cells were treated with CM from HCC cells with different expression levels of CLIC1. (C) The number of EA.hy926 cells was measured using ImageJ software.
Supplier Page from Proteintech Group Inc for CLIC1 antibody