Fig. 1: USP9X is highly correlated with TGF-β signaling during HGOSC progression and chemoresistance. | Cell Death & Disease

Fig. 1: USP9X is highly correlated with TGF-β signaling during HGOSC progression and chemoresistance.

From: USP9X integrates TGF-β and hypoxia signalings to promote ovarian cancer chemoresistance via HIF-2α-maintained stemness

Fig. 1

A Representative images of p-Smad3 and USP9X IHC stainings on differently progressed HGSOC tissues (n = 83), including benign (n = 17), borderline (n = 25), and malignant tumors (n = 41). Scale bars, 50 µm. The images under are the zoom-in images for the upper images in each group. The right panels were clinical scores, and correlation analysis between p-Smad3 and USP9X. B Immunoblotting (IB) analysis of p-Smad3 and USP9X on freshly collected tumor tissues of serous OC with varying progression, including benign (n = 9), borderline (n = 9), and malignant tumors (n = 9). C GSEA of TGF-β signaling pathway in chemotherapy or platinum-resistant patients based on GSE241221 (scRNA-sequencing), GSE51373 and GSE131978. D IB analysis of p-Smad3, Smad2/3 and USP9X in ID8, chemoresistant cells ID8-rCDDP or ID8-rPTX with or without short treatment (24 h) of CDDP (10 μM) or PTX (5 nM). E Representative images of E-Cadherin, N-Cadherin, Twist, and ALDH1A1 IHC stainings on different progress HGSOC tissues (n = 83), including benign (n = 17), borderline (n = 25), and malignant tumors (n = 41). Scale bars, 50 µm. The right panels were clinical scores. F Representative images of spheroids in fresh HGOSC samples (n = 32). Spheroids larger than 50 μm in diameter were used for analysis. Scale bars, 75 µm. G Correlation analysis between spheroid numbers and p-Smad3 or USP9X expression based on (F). H GSEA analysis of TGF-β signaling pathway between USP9X high and low groups based on TCGA. Smad2 binding motif (I) and potential SREs on USP9X promoter (J) based on Jasper database. K Dual-luciferase assays were performed to analyze the transcriptional activation of USP9X promoter with TGF-β1 (5 ng/mL), CDDP (10 μM) or PTX (5 nM) for 24 h in CAOV3 cells. ChIP-qPCR was used to detect the Smad2/3 binding onto −1121 bp site of USP9X promoter, the % Input data was shown in (L) and the agarose gelatin plot was shown in (M). Data are shown as the mean ± s.d (A, E, K, L). P values were calculated by unpaired two-tailed Student’s t test (A, E, K, L). Correlation analysis was performed by Pearson correlation, P values (two-tailed) were calculated by Pearson r (A, G). n = 3 biological independent samples (D, K, L, M).

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