Fig. 7: Role of TRPM7 kinase domain in KrasG12C PDAC cell line. | Cell Death & Disease

Fig. 7: Role of TRPM7 kinase domain in KrasG12C PDAC cell line.

From: The kinase domain of TRPM7 interacts with PAK1 and regulates pancreatic cancer cell epithelial-to-mesenchymal transition

Fig. 7

A Representative morphology of MIA PaCa-2 Control and ΔK cells at ×100 magnification. Scale bar=200 µM. B Circularity index calculated for MIA PaCa-2 Control (n = 71) and ΔK (n = 98) cells (p < 0.001, Mann–Whitney test). C Cell migration and invasion of MIA PaCa-2 Control and ΔK evaluated in Boyden chambers (n = 3, p < 0.001, Mann–Whitney test). Scale bar=200 µM. D mRNA expression of the RhoGTPase family Pak1, Rac1, Rac2, Rac3, RhoA in MIA PaCa-2 Control and ΔK cells (n = 8, p < 0.05 and p < 0.01; Two-way ANOVA followed by Šidák’s post-hoc test). E Protein expression levels of pPAK1 (n = 4), PAK1 (n = 8, p < 0.05, Mann–Whitney test), RAC (n = 6, p < 0.01, Mann–Whitney test) and RhoA (n = 6, Mann–Whitney test in MIA PaCa-2 Control and ΔK cells. F Co-immunoprecipitation of TRPM7 with PAK1 in MIA PaCa-2 Control and ΔK cells and their respective positive and negative controls. G Proximity Ligation assay of TRPM7 with PAK1 in MIA PaCa-2 cells incubated with DMSO (n = 272) or 50 µM TG100-115 (n = 277) for 24 h (Mann–Whitney test). H Cell viability of MIA PaCa-2 Control and ΔK cells assessed by MTT assay during 96 h (n = 3, p < 0.001, Two-way ANOVA followed by Šidák’s post-hoc test). I Cycle cell distribution of MIA PaCa-2 Control and ΔK evaluated by flow cytometry (n = 3, p < 0.01 and p < 0.001, Two-way ANOVA followed by Šidák’s post-hoc test). J Expression and phosphorylation levels of CYCB1 and CDK1 proteins in MIA PaCa-2 Control and ΔK cells (n = 8, p < 0.05 and p < 0.01, Two-way ANOVA followed by Šidák’s post-hoc test). K Monitoring of tumor volume on SCID mice injected with MIA PaCa-2 Control (n = 6) and ΔK (n = 6) for 54 days (Two-way ANOVA). L Masses in grams of tumors weighted after mice sacrifice (n = 6, Student t-test). M Detection of micrometastasis in the liver and lungs of mice injected with MIA PaCa-2 Control and ΔK cells (Fisher test). Human GAPDH was used to detect micrometastasis in those organs. Results are shown as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001.

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