Fig 1: ZNF711 regulates SLC31A1 promoter activity in EOC cells. (a) The SLC31A1 promoter luciferase reporter plasmids and Renilla pRL-TK plasmids were transfected into the indicated empty vector-transduced, ZNF711-transduced, control shRNA-transduced, and the ZNF711-shRNA transduced HEY and SKOV3 cells. After 48 h, the luciferase signal was examined and presented. (b) Schematic illustration of the P1-P7 cloned fragments of the human SLC31A1 promoter. (c and d) Luciferase activity of the SLC31A1 promoter fragments in control vector cells and ZNF711-overexpressing EOC cells. (e and f). Luciferase activity of SLC31A1 promoter fragments in shRNA control vector cells and ZNF711 knockdown EOC cells. (g) ChIP assays were performed using an anti-Flag antibody to screen three predicted ZNF711-binding sites on the SLC31A1 promoter. IgG was used as a negative control. (h) ChIP assay showing the enrichment of Pol II on region 2 in the SLC31A1 promoter in the indicated HEY and SKOV3 cells. Each bar shown in the figure represents the mean ± SD of three independent experiments. *p < 0.05 (unpaired t-test or one-way ANOVA with Bonferroni's correction).
Fig 2: The effects of oxaliplatin (OX) with or without ß-elemene (ELE) on in vivo tumor growth in an orthotopic implantation mouse model of MHCC97H cells and on CTR1 protein expression in tumor tissues.Oxaliplatin with ß-elemene suppressed tumor growth in orthotopic implantation mouse models of MHCC97H cells. (a) Tumor volumes and weight in the orthotopic implantation model mice at week 7. (b) The weight in the orthotopic implantation model mice at the indicated time points. (***p < 0.001) (c) Representative examples of immunohistochemistry staining for CTR1 protein from MHCC97H xenografts (400 × magnification; scale bars, 50 µm). (d) Representative examples of tumor xenografts stained with apoptosis by TUNEL assay (positive cell labelled with arrows; 400 × magnification; scale bars, 50 µm).
Fig 3: Effects of reduced intracellular copper by knocking-down CTR1 or TEPA copper chelation on HMEC-1 angiogenic activity. (a) Representative western blot for CTR-1 protein (35 kDa cropped band) on whole-cell extracts from HMEC-1 cells. GAPDH (37 kDa cropped band) expression was used as a protein loading control. (b) Representative photographs of HMEC-1 cells in Matrigel™ assays. Photographs were taken after knockdown of CTR-1 using 20 nM CTR-1 siRNA B or 8 h incubation with TEPA. Non-silencing siRNA (NSsiRNA) was use as control. Inset, % of inhibition as compared to untreated control cells; N.S. non-significant; Scale bar 200 µM. (c) Total surface area of vascular structure. Columns, n = 3, bars, SEM (****p < 0.0001).
Fig 4: Knockdown of CTR1 in IMR-32 and BE(2)-C cells significantly reduced their sensitivity to Dextran-Catechin(A) Cell death in IMR-32 and (B) BE(2)-C cells after knockdown of CTR1 and subsequent treatment with Dextran-Catechin for 24 hours. Columns, means of at least three independent experiments; Bars, SEM (***p < 0.001, ****p < 0.0001).
Fig 5: Combined BIX-01294 sensitizes ZNF711-downregulated EOC cells to CDDP therapy. (a-b) SLC31A1 mRNA (a) and protein expression (b) in ZNF711- down regulated EOC cells treated with BIX-01294 or vehicle control. (c) MTT analysis (left) and IC50 value (right) of CDDP in the indicated cells. (d) Representative images (left) and quantification (right) of the number of colonies in the indicated cells treated with either the vehicle or BIX-01294. (e) FACS analysis of Annexin V staining (left) and quantification (right) of indicated stable HEY and SKOV3 cell lines treated with a vehicle or BIX-01294. (f) Intercellular CDDP content in the indicated cells. (g) The quantification of DNA-bound CDDP (Pt) in the indicated cells. (h) Representative images of the indicated nude mice treated with a vehicle or BIX-01294. (i) Relative change in the luminescence signals of the indicated xenograft tumors at the indicated time points. (j) Kaplan-Meier survival analysis of the indicated tumor-bearing mice. (k) Concentration of intercellular CDDP in indicated cells. (l) IHC staining of SLC31A1, PCNA, and TUNEL staining (left) and quantification (right) of SLC31A1 expression, apoptotic rate, and proliferation index in the indicated xenograft tumors. Scale bars, 50 µm. Each bar shown in the figure represents the mean ± SD of three independent experiments. *p < 0.05 (one-way ANOVA with Bonferroni's correction).
Supplier Page from Abcam for Anti-SLC31A1 / CTR1 antibody [EPR7936]