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Fig. 2 | Molecular Cancer

Fig. 2

From: Circular RNA MTCL1 promotes advanced laryngeal squamous cell carcinoma progression by inhibiting C1QBP ubiquitin degradation and mediating beta-catenin activation

Fig. 2

The effect of circMTCL1 on LSCC biological functions by regulating C1QBP ubiquitination. a, Silver staining images of PAGE gels, in which circMTCL1/proteins complexs from pull down experiments of TU212 cells were separated; the arrows represent the specific protein bands in pull down complexs by circMTCL1 sense sequence when compared with antisense sequence. b, Western blot assays for SDS-PAGE electrophoresis of RNA pull-down assay. c, The peak map of C1QBP acquired from the RNA pulldown mass spectrometry assay. d, KEGG analysis of the protein C1QBP, and the related signaling pathways. e, Western blot assays showing the effect of circMTCL1 on the proteins pulled down by target RNA probe (circMTCL1). f, The C1QBP expression was conducted after overexpressing or knocking down circMTCL1 by Western blot. g, CircMTCL1 expression was determined after overexpressing, knocking down C1QBP by qRT-PCR assays . Values are the mean ± s.d. of n = 3 independent experiments. h, Reducing the translation activity of C1QBP by silencing circMTCL1 treated by the CHX in TU212 cells (left). The quantification of C1QBP degradation rate by gray scale analysis (right). i, Western blot showing C1QBP protein in TU212 cells after knocking down circMTCL1 and then treated with MG132 for 24 h. j, Ubiquitinated C1QBP measured by immunoprecipitation with anti-C1QBP antibody or IgG control and immunoblotting with anti-ubiquitin antibody in TU212 cells after knocking down circMTCL1. k, IHC method was used to determine the cell localization and the expression levels of C1QBP in LSCC tissues and matched adjacent-tumor controls (n = 140). Scale bars = 100 μm. l, Violin charts illustrating the expression scores and levels of C1QBP between LSCC tissues and corresponding adjacent normal tissues (n = 140). Nonparametric tests and Median and 95%CI were shown. m, Kaplan-Meier curves displaying the impact of C1QBP on overall survival in the included LSCC cohort. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001

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