Figure 7:

CRISPR-Cas9 mediated deletion of Rtl3 impairs chondrogenesis and delays matrix synthesis in ATDC5 cells. (A) ATDC5^Rlt3−/− cells did not express Rtl3 mRNA at any chondrogenic time point. (B) Reduced matrix deposition was visualized in wild type and ATDC5^Rlt3−/− cells with Alcian Blue stain (scale bar 100 μm) and (C) semi-quantified by percent area analyses (ImageJ). (D) Sox-9 mRNA levels were significantly reduced in ATDC5^Rlt3−/− cells assessed by qPCR. ATDC5^Rlt3−/− cells demonstrated a significant reduction in Col2a1 mRNA (E) and (F) protein levels compared to wild type ATDC5 cells.