Fig. 2.

ER stress increases the lysosomal level of CMA regulator LAMP2A. a ER stress increases the level of CMA regulator LAMP2A. Total or purified lysosomal lysates from SN4741 cells treated with Tg or Tu for 0–12 h were blotted as indicated. b ER stress increases LAMP2A on lysosomal membrane. Whole lysosomes, lysosomal membrane, and matrix were prepared from SN4741 cells treated with Tg for 12 h and blotted for LAMP2A and Hsc70. c ER stress induces LAMP2A oligomerization. Lysosomes purified from Tg-treated SN4741 cells were subjected to native continuous gel electrophoresis and immunoblotted for LAMP2A. d ER stress does not affect the level of LAMP2A mRNA. SN4741 cells were treated with Tg (0.3 μM) for different time periods. The levels of LAMP2A mRNA were determined by qRT-PCR. Data are shown as mean ± s.d. n = 3 × 4 repeats (One-way ANOVA with Turkey, p > 0.05). Bottom panels of a and b show quantification of LAMP2A and LAMP2A/Hsc70 relative to basal condition (n = 3 of independent experiments). All values are mean ± s.d. (ANOVA with Turkey test for a, unpaired two-tailed t test for b; *p < 0.05 or **p < 0.005 vs. control)