FIGURE 4.

TFEB-regulated lysosomal genes and TFEB levels are unaltered in acute TRPML1 and PPT1 KD cells. A, mRNA levels were measured in TRPML1 and PPT1 KD cells using qRT-PCR using CatB, LAL, and LAMP-1 specific primers (n ≥ 3). No significant change was seen in mRNA levels under either condition. B, TFEB protein levels were measured by Western blot analysis in TRPML1 KD cells. No difference was seen. C, TFEB cellular localization was analyzed by confocal microscopy. No change in localization occurs after 48 h of TRPML1 KD (n = 3). D, qRT-PCR analysis of CatB, LAL, and LAMP1 mRNA in HeLa cells exposed to 100 mm sucrose for 2 or 5 days to induce general inhibition of the lysosomal function as described before. E, qRT-PCR analysis of the same genes in cells after a 7-day KD of TRPML1 or PPT1. β-Actin was used as a loading control (*, p < 0.05, n = 3).