Figure 6
From: Loss of chaperone-mediated autophagy is associated with low vertebral cancellous bone mass
CMA deficiency increases RANKL expression and decreases mineral deposition of an osteoblastic cell line. Lamp2a was knocked down in osteoblastic UAMS-32 cells and plated into 6 well plates for gene expression or mineralization assays. (a–c) qRT-PCR analysis of three Lamp2 isoforms and Tnfsfs11 (Rankl) in osteoblastic UAMS-32 cells that stably express scrambled shRNA (shRNA Con) or shRNA targeting LAMP2A (shRNA L2A). mRNA levels of all transcripts were normalized to the housekeeping gene β-actin. n = 6 wells/group. Bars indicate mean ± STDEV. p values were calculated and evaluated by Welch's t-test; *, p < 0.05. Knockdown of Lamp2a and the following gene expression was performed an additional time independently and shown in S. Fig. 6a–c. (d) shRNA Con and shRNA L2A cells were cultured in osteogenic media. Mineral deposition was measured by Alizarin Red staining on day 4 and day 8 of culture.
