KD of CTGF expression inhibits proliferation of MSCs isolated from BM. (A) CTGF knockout or control pups were killed, and their stroma-rich body organs, including liver, thymus, spleen, and BM, were surgically dissected. Cell suspensions of these organs were cultured in α-minimum essential medium containing 20% fetal bovine serum fibroblast-like colony-forming units and were observed 7 to 10 days after plating in controls but not in CTGF knockout cells. (B) Normal MSCs derived from human BM were transduced with a lentivirus expressing CTGF shRNA. Data represent relative expression of CTGF to GAPDH in control and CTGF KD MSCs. (C) CTGF protein expression was analyzed in cell lysates from control and CTGF KD MSCs by western blotting with anti-CTGF antibody. α-Tubulin served as a loading control. (D) Morphology of control and CTGF KD MSCs. (E) Cell proliferation was analyzed by counting absolute cell numbers with a Vi-Cell XR cell counter. Control or CTGF KD MSCs (2 × 10⁴) were cultured in 6-well dishes in triplicate and counted on days 2, 3, 4, and 5.