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. 2019 Oct 17;13(1):45–60. doi: 10.1007/s12195-019-00602-2

Figure 4.

Figure 4

CCN4 promotes EMT gene expression in mouse B16 melanoma cells. (a) Protein expression comparison of some EMT-associated genes in B16F10 and its knockout cells. Βeta-actin was used as loading control for western blotting. (b) mRNA expression comparison, as revealed by real-time quantitative RT-PCR, of some EMT-associated transcription factors and EMT marker genes in B16F10 and its knockout cells. (c–f) mRNA expression change of EMT-associated transcription factors and EMT marker genes in B16F10 knockout cells after CCN4 was supplied in culture medium. Both B16F10 and B16F10-KO cells were plated in complete DMEM with 10% FBS for 48 h before recombinant mouse CCN4 protein (rmCCN4) was added to B16F10-KO cells (5 µg/mL), or B16F10 medium was transferred to B16F10 –KO cells. Cells at indicated time points were harvested for RNA isolation and real-time quantitative RT-PCR. **p value < 0.01; ***p value < 0.001.