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. 2021 Jan 1;24:528–541. doi: 10.1016/j.omtn.2020.12.023

Figure 5.

Figure 5

USP37 promotes the expression of CDT1 by deubiquitination

(A) Differential expressed genes in the dataset obtained from the GEO database (GEO: GSE106929) represented in a volcano plot (red dots indicate upregulated genes and green dots indicate downregulated genes in lung tumor samples). (B) Differentially expressed genes in the dataset obtained from the GEO database (GEO: GSE33532) represented in a volcano plot (red dots indicate upregulated genes and green dots indicate downregulated genes in lung tumor samples). (C) Overlapped upregulated genes in datasets GEO: GSE106929 and GSE33532 visualized by a Venn diagram. (D) Expression levels of USP37 and CDT1 were co-related by an online tool (Chipbase). (E) Relative expression levels of CDT1 in lung tumor (red box) and normal lung tissue (gray box) analyzed by TCGA database. (F) Representative images and results of immunoblotting analysis evaluating CDT1 expression in lung tissues obtained from lung cancer patients with OSAH (n = 27) or without OSAH (n = 68) (∗p < 0.05, lung cancer + OSAH versus lung cancer). (G) Correlation analysis between miR-320b and CDT1 as well as between USP37 and CDT1 in lung cancer patients with OSAH. (H) Representative images and results of immunoblotting analysis evaluating CDT1 expression in normal bronchial epithelial cells and lung cancer cells (∗p < 0.05, lung cancer cells versus NHBE cells). (I) Representative images and results of immunoblotting analysis validating the overexpression or knockdown of USP37 (∗p < 0.05, oe-USP37 versus oe-NC; #p < 0.05, sh-USP37 versus sh-NC). (J) Representative images and results of immunoblotting analysis evaluating CDT1 expression in lung cancer cells with the overexpression or knockdown of USP37 (∗p < 0.05, oe-USP37 versus oe-NC; #p < 0.05, sh-USP37 versus sh-NC). (K) Representative images of co-immunoprecipitation assay investigating the interactions between USP37 and CDT1. (L) Representative images of immunofluorescent staining showing co-localization of USP37 and CDT1 (magnification 400×). (M) Representative images of immunoblotting analysis evaluating the expressions of USP37 and CDT1. (N) Ubiquitination assay detecting ubiquitination levels of USP37 and CDT1. Data are presented as the mean ± standard deviation. Data between two groups were compared using an unpaired t test. Comparisons among multiple groups were performed using one-way ANOVA with Tukey’s post hoc test. The correlation between two indicators was analyzed by Pearson correlation coefficient.