Fig 2: Levels of TPC1 and TPC2 mRNA relative to ß-actin mRNA (A). Western-blot of TPC1 and TPC2 proteins (B) isolated from rat aorta SMCs and relative levels of TPC1 and TPC2 (C). The values panel A are the means of four measurements in different SMCs preparation + SEM (* p < 0.01). In panel B, the typical image of Western blots from one of three different cell lysates is presented. (+) Marks hybridization in the presence of blocking peptide. In panel C, n = 3 and n.s.—not significant.

Fig 3: Effect of PM0.1 and NP20 on expression and activity of the lysosomal channel TRPML1. (A) Superimposed representative traces (a) and quantification (b) of the effect of ML-SA1 (10 µM) on [Ca2+]i in NSC-34 motor neurons exposed to PM0.1 (2.86 ppm) and NP20 (0.71 ppm) (48 h) in the presence or absence of Trolox (100 µM). This latter drug was added every 24 h. All the experiments were repeated three times on almost 20 cells for Control, Control+Trolox, Control+PM0.1, Control +NP20 and 25 cells for PM0.1+Trolox and NP20+Trolox. * p < 0.05 vs. Control; ** p < 0.01 vs. PM0.1 or NP20. (B) Representative Western blotting of TRPML1 expression in NSC-34 motor neurons exposed to PM0.1 and NP20 (48 h) (a) and quantification (b). All the experiments were repeated three times on different preparations. * p < 0.05 vs. control. (C) Representative Western blotting of TPC2 expression in NSC-34 motor neurons exposed to PM0.1 and NP20 (48 hrs) (a) and quantification (b). All the experiments were repeated three times on different preparations.