Fig. 7

TMEM106B regulates the degradation of LAMP2A by influencing the interaction between LAMP2A and CTSA. (A) LAMP2A degradation with or without overexpression of TMEM106B in HeLa cells. HeLa cells were transfected with HA-LAMP2A with or without GFP-TMEM106B, treated with 10 μg/mL cycloheximide (CHX) for specified times, and used for Western blot analysis. Overexpression of TMEM106B slowed down the degradation of LAMP2A. (B) Western blot analysis showed that the expression of mature-CTSA was not affected by knockdown of TMEM106B with siRNA compared with NC siRNA in HeLa cells (n = 3 samples per group). (C) Co-IP analysis showing the interaction between CTSA and LAMP2A in HeLa cells with co-expression of FLAG-CTSA and HA-LAMP2A. (D) Co-IP analysis showing the interaction of CTSA and LAMP2A with or without the overexpression of TMEM106B. The graph on the right shows quantification of the binding of CTSA to LAMP2A (n = 4 samples per group). *p < .05, **p < .01, ns, no significance; Data were shown as mean ± SD; Statistical analysis was performed using a Shapiro–Wilk test for samples with normal distribution and paired student's t tests with two tails.