Fig. 5: CircTICRR interacting with HuR stabilizes GLUD1 mRNA. | Cell Death & Disease

Fig. 5: CircTICRR interacting with HuR stabilizes GLUD1 mRNA.

From: Oncogenic circTICRR suppresses autophagy via binding to HuR protein and stabilizing GLUD1 mRNA in cervical cancer

Fig. 5

A The binding of circTICRR to the AGO2 protein was determined by agarose gel electrophoresis assay. B Left panel: RNA pull-down assay of SiHa cells using a junction-specific circTICRR probe or an oligo probe. Sliver staining and MS then were performed to analyze the pulled down differentially expressed proteins. Right panel: Venn diagram showing the overlap results of the differentially expressed proteins in MS and circTICRR targets in StarBase. C, D. Protein pulled down by control oligo or circTICRR probe with HuR antibody was detected by western blot (C), and enriched circTICRR using HuR antibody was detected by qRT-PCR after RIP assay (D) of SiHa and CaSki cells. Data were calculated as input %. E, F. Protein pulled down by control oligo or GLUD1 mRNA probe with HuR antibody was detected by western blot (E) and enriched GLUD1 mRNA using HuR antibody was detected by qRT-PCR after RIP assay (F) of SiHa and CaSki cells. Data were calculated as input %. The relative levels of GLUD1 mRNA (G) and protein H expression in the SiHa and CaSki cells with HuR knockdown were detected by qRT-PCR and western blot, respectively. I SiHa and CaSki cells transfected with a negative control siRNA or two HuR siRNAs were treated with actinomycin D (5 μg/ml) at the indicated time points. The relative level of GLUD1 mRNA expression was detected by qRT-PCR. J SiHa and CaSki cells transfected with a negative control siRNA or two circTICRR siRNAs were treated with actinomycin D (5 μg/ml) at the indicated time points. The relative GLUD1 mRNA level was detected by qRT-PCR. The relative level of HuR expression in the SiHa and CaSki cells with circTICRR knockdown was detected by western blot (K) and qRT-PCR L, respectively. M. GLUD1 mRNA recruited by HuR antibody from the lysates in SiHa cells with or without circTICRR knockdown was detected by RIP assay followed by qRT-PCR. Data are representative of at least three independent experiments and presented as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001.

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