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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: LncRNA SPOCD1-AS from ovarian cancer extracellular vesicles remodels mesothelial cells to promote peritoneal metastasis via interacting with G3BP1

Fig. 6

SPOCD1-AS induces the MMT process via binding with G3BP1. a RNA pull-down assays in MeT-5A cells using biotin-labeled sense and antisense SPOCD1-AS probes. Sliver staining and mass spectrometry were performed to analyze the proteins pulled down. The black arrow indicates G3BP1 protein band. b Western blot validation of protein pulled-down using biotin-labeled sense and antisense SPOCD1-AS probes with G3BP1 antibody. GAPDH was used as the loading control. c RNA immunoprecipitation assays in MeT-5A cells using G3BP1 antibody to enrich SPOCD1-AS. GAPDH and U1 were used as negative controls. Data were calculated as input%. d RNA-FISH assays of SPOCD1-AS (red) and immunofluorescence of G3BP1 protein (green) in MeT-5A cells. Scale bar, 10 μm. e, f, g MeT-5A cells were transfected with two G3BP1 siRNAs and negative control, respectively, for 48 h. MMT-related proteins were detected by western blot analysis (e), migration and adhesion assays were performed. Scale bar, 100 μm (f, g). h, i, j MeT-5A cells were transfected with SPOCD1-AS lentivirus, SPOCD1-AS lentivirus plus si-G3BP1#1, SPOCD1-AS lentivirus plus si-G3BP1#2, and negative control, respectively. MMT-related proteins were detected by western blot analysis (h), migration and adhesion assays were performed. Scale bar, 100 μm (i, j). Data are representative of at least three independent experiments and are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001

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