Research Paper Volume 14, Issue 12 pp 5250—5270

Figure 2. Expression profiles of TRPM7 in HNSCC-derived cell lines and HNSCC tumor samples as well as knockdown of TRPM7/NFAT expression via shRNA. (A) Immunoblotting analysis of TRPM7 in HNSCC-derived cell lines and NHOKs, quantification of TRPM7 mRNA expression in HNSCC-derived cell lines through qRT-PCR, and target expression screening strategy for head and neck cancer cell lines. (B) qRT-PCR and protein analysis of SAS cells with TRPM7 knockdown and control cells in the TRPM7/NFAT axis. Knocking down of TRPM7 downregulated the expression of genes and proteins related to TRPM7 and the calcineurin/NFAT pathway (NFATC3, NOTCH1), which implies that TRPM7 and the calcineurin/NFAT pathway may have a regulatory relationship, as well as representative images of the SAS cell colony and sphere (C, D). (E) Bar graph shows the mean ± standard deviation of the percentage of migrating and invasive SAS cells treated with shNC or shTRPM7 over three independent experiments. (F) Western blot analysis results showed target of TRPM7 that may affect invasion and migration pathway of HNSCC by evaluating modulation of several EMT markers. (G) Analysis of the relationships of TRPM7 with E-caderin and vimentin. *p < 0.05, **p < 0.01.