CTGF KD MSCs differentiate into adipocytes in vitro and in vivo. (A) To examine the differentiation potential of control or CTGF KD MSCs, cells (5 × 10⁴) were cultured in adipocyte differentiation medium for 28 days. After incubation, the cells were stained by Oil Red O dye or LipidTox fluorescent dye to observe adipocyte differentiation. (B) Quantitative representation of data showed in supplemental Figure 4. (C) To examine the differentiation potential of CTGF KD MSCs in vivo, a model of EXM-BM was developed by injecting human MSCs (1.5 × 10⁶) mixed with human endothelial progenitor cells (1.5 × 10⁶) in 0.2 mL Matrigel subcutaneously into the flanks of NOD/SCID/IL-2rγ^null mice. Control cells (empty vector and nonspecific shRNA controls) were transplanted on the left, and CTGF KD MSCs were transplanted on the right. (D) Eight weeks after transplantation, the bone pellets were dissected and fixed in 4% paraformaldehyde. Tissue sections were then stained with hematoxylin and eosin to observe tissue architecture. Scale bar represents 200 μm. (E) To investigate adipocyte differentiation, immunohistochemical analysis was performed on EXM-BM using a PPARγ or cEBPα antibody.