Fig 2: Wisp2 was required for BMSC migration and differentiation to endothelial cellsSignificantly decreased expression of Wisp2 and Cxcr4 in rat livers after the Wisp2-sgRNA-BMSC transplanted 18 days, the expression of Wisp2 (a) and Cxcr4 (b) were determined in liver sections. Liver cyrosections were stained for GFP (Green fluorescence) and Wisp2/Cxcr4 (Red fluorescence). Images were merged with DAPI (Blue fluorescence) staining reveal the nuclei. (c) Expression of the endothelial cell marker CD31 (Red fluorescence) was also examined in the liver sections by immunostaining. The expression of CD31 was reduced in the Wisp2-sgRNA BMSCs transplanted rat livers.

Fig 3: High expression of Wisp2 in BMSCsTotal RNA was extracted from rat BMSCs or rat livers, then PCR (a) and qRT-PCR (b) were performed to examine Wisp2 mRNA level. Error bars represent the ± S.E.M. of triplications. “***”, p<0.001. Gene expression level of the samples normalized to Gapdh and the expression of Wisp2 in liver was set as 1. (c) The protein level of Wisp2 was measured in BMSCs and livers with western blot analysis. 20µg total protein per lane and Gapdh was used as an internal control.

Fig 4: Cxcr4 was decreased and repair capability was reduced in lentiviral Wisp2 disrupted BMSC transplantation ratsLiver samples treated with Wisp2-sgRNA or not were examined for the expression of Cxcr4 (a) and IL-6 (b). 40μg of total protein per lane, as indicated by western analysis 18 days after transplantation. (c) Paraffin- embedded sections from livers obtained 18 days after the transplantation were stained with H&E and PCNA or α-SMA. The Wisp2-sgRNA group showed more extensive liver damage and cellular swelling (20×). Serum ALT (d) and AST (e) levels were assessed to determine the degree of liver injury. The control group was set as 1. Asterisks indicate significant differences, “*”, P< 0.05. (f) Representative micrographs of TUNEL staining of liver tissue. Liver sections collected 18 days after BMSCs transplantation were stained with FITC-conjugated TUNEL to identify apoptotic cells. The results showed that disrupted Wisp2 incresed the levels of apoptosis caused by 2-AAF/PH. Original magnification: 20×.

Fig 5: Wisp2 was disrupted in rat livers after 2-AAF/PH(a) Seven days after the intragastric administration of 2-AAF, the rats underwent PH and were transplanted with 1.8×106 Wisp2-sgRNA-BMSCs or equal amounts of control BMSCs and then sacrificed and livers collected as indicated in the scheme. (b) Representative images showing homing BMSCs to the injured liver. (c and d) Wisp2 expressed in rat livers. Expression of Wisp2 was detected in rat livers 18 days post-transplantation by Immunoblot analysis (c). 40µg of total protein per lane. (d) Immunostaining was performed. Original magnification: 10× (Left panel) and 40× (right panel).