Fig. 5: TUBB3 is the functional downstream target of RPPH1 in CRC cells. | Cell Death & Disease

Fig. 5: TUBB3 is the functional downstream target of RPPH1 in CRC cells.

From: LncRNA RPPH1 promotes colorectal cancer metastasis by interacting with TUBB3 and by promoting exosomes-mediated macrophage M2 polarization

Fig. 5

a Relative TUBB3 mRNA levels were quantified by qRT-PCR in SW620 cells with stable RPPH1 overexpression or knockdown. 18S rRNA served as the control. b WB analysis for TUBB3 protein levels in stable RPPH1 overexpression or knockdown SW620 and HCT8 cells. c, d SW620 cells with stable RPPH1 overexpression or knockdown were treated with cycloheximide (CHX, 50 μg/ml) for the indicated times and TUBB3 protein levels were analyzed via WB analysis. e Stable RPPH1 knockdown SW620 and HCT8 cells were treated with MG132 (25 μM) for 12 h and then TUBB3 protein levels were analyzed via WB analysis. f Cell lysates form stable RPPH1 overexpression or knockdown SW620 cells treated with MG132 for 12 h were immunoprecipitated (IP) with either control IgG or TUBB3 antibody and then immunoblotted for ubiquitin and TUBB3. g WB analysis for TUBB3 protein levels in stable TUBB3 overexpression or knockdown SW620 and HCT8 cells. h, i Transwell migration and invasion assays in stable TUBB3 overexpression (h) or knockdown (i) HCT8 cells. j The EMT effect was validated by WB analysis of epithelial or mesenchymal markers in SW620 cells. k Rescue assays for WB analysis of the change of EMT markers were performed in SW620 cells with RPPH1 and TUBB3 changing. Values are represented as mean ± SD. NS no significant. **p < 0.01, and ***p < 0.001

Back to article page