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. 2021 Apr 19;11:578928. doi: 10.3389/fonc.2021.578928

Figure 1.

Figure 1

Transient receptor potential mucolipin 1 (TRPML1) silencing affects glioblastoma cell viability. (A) The relative TRPML1 mRNA expression in untransfected and siTRPML1 T98 and U251 glioma cell lines after 72 h of silencing (time 0) and after 72-h post-transfection was evaluated by the quantitative reverse transcription (qRT)-PCR. TRPML1 mRNA levels were normalized for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression. Data are expressed as mean ± SD; *p < 0.001 vs. untransfected. (B) The Western blot analysis of TRPML1 protein levels. Blots are representative of one of three separate experiments. TRPML1 densitometry values were normalized to GAPDH used as loading control. The TRPML1 protein levels of silenced samples were determined with respect to TRPML1 levels of untransfected cells. (C) The Western blot analysis of pERK, ERK, pAKT, and AKT protein levels in siGLO and siTRPML1 T98 and U251 cells after 72 h of silencing (time 0). Blots are representative of one of three separate experiments. ERK and AKT densitometry values were normalized to GAPDH used as loading control. The pERK and pAKT protein levels were determined with respect to ERK and AKT levels. *p < 0.01 siTRPML1 vs. siGLO. (D) Cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in siGLO control and in siTRPML1 T98 and U251 glioblastoma cells for up to 72 h. Data shown are expressed as mean ± SE of three separate experiments. *p < 0.01 and #p < 0.05 siTRPML1 vs. siGLO.