Fig 1: STC1 promotes metastasis and lipid metabolism in vitro. A WB results of the STC1 expression levels in different OC cell lines. B STC1-knockdown cell lines were established with Skov3-ip1 and Hey cells, and the transfection efficiency was confirmed by WB assays. C PCA of data from untargeted relative quantitative lipidomic analysis. D Volcano plot of individual lipid species significantly up- or down- regulated in the NC group versus the STC1-sh1 and STC1-sh2 groups in Hey cell lines (lipids with a P value < 0.05 and a fold change (FC) > 1.5 or < 0.67 were indicated in blue or red, respectively). E Correlation analysis of the ten lipid classes. F A heat map of significant differences in the variables among the NC, STC1-sh1 and STC1-sh2 groups of Hey cells. The variables were ranked by ANOVA. G TEM images of OC cells in Skov3-ip1 and Hey cell lines (red arrow: mitochondria). H ORO staining of OC cells in Skov3-ip1 and Hey cell lines (red arrow: TGs). (Data are shown as the mean ± SD values. Significance was calculated using Student’s t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.)
Fig 2: STC1 promotes metastasis, lipid metabolism and DDP chemoresistance in vivo. A The model of cell injection into nude mice. B-C Luciferase expression was detected in the mouse model. The NC group showed significantly higher flux than the NC + DDP group, STC1-sh1 group and STC1-sh1 + DDP group. D The NC group showed significantly larger tumors than the NC + DDP group, STC1-sh1 group and STC1-sh1 + DDP group. E The tumor weights and tumor volumes in the four groups. F-G The NC groups had metastatic tissues in the liver, spleen and intestine. H H&E staining of metastatic tissues (liver, spleen and intestine) in the NC groups. I Representative WB results of UCP1, TOM20 and perilipin1 in tissues from two mouse tumors generated by injection of the two cell lines. J IF analysis showed the relationship between STC1 and UCP1 in tissues from two mouse tumors generated by injection of the cell lines. (Data are shown as the mean ± SD values. Significance was calculated using Student’s t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.)
Fig 3: Association between STC1 and CD3+ T cells infiltration level. (A) Representative IHC images showing BLCA samples immunostained for STC1 and CD3. (B) Different density of CD3+ T cells infiltration according to STC1 levels. *P value < 0.05.
Fig 4: Western blot analysis. (A) The correlation between STC1 and TGFß1 was analyzed. (B) The expression levels of STC1, P-SMAD2/3 and SMAD4 were detected via western blotting. *P<0.05 vs. respective LV3-NC group. STC1, stanniocalcin-1; TGFß1, tumor growth factor ß1. TPM, transcripts per million.
Fig 5: miR-34a regulates the expression of SMAD4 and STC1. (A) Silencing STC1 affected the expression levels of various miRNAs in LN229 cells. (B) Overexpression of STC1 regulated the expression level of STC1 in T98G cells. (C) Effect of miR-34a mimics and inhibitors. (D and E) miR-34a regulated the expression of STC1 and SMAD4. (F) A dual luciferase reporter gene assay was performed. Error bars represent the standard error. *P<0.05. miRNA, microRNA; miR, microRNA; STC1, stanniocalcin-1; NC, negative control; 3'UTR, 3' untranslated region.
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