Fig 2: Hypoxia induces EMT and VM formation in glioblastoma cells(A) Immunofluorescence was analyzed for MIF, CXCR4 and endothelial marker (E-cadherin) in U87and U251cells exposed to hypoxia. Images obtained at magnification 200X. Relative mRNA expression levels of MIF and CXCR4 were detected by Q-PCR. (B) Western blots showed the expression levels of MIF, CXCR4 and multiple markers (E-cadherin, N-cadherin, Vimentin) associated with EMT in U87 and U251 cells under hypoxic conditions. Relative mRNA expression levels of E-cadherin and Vimentin were detected by Q-PCR. The data shown are the mean ± SD of independent experiments, n = 3. (C) VM tube formation assay was performed 6h with U87 cells under hypoxia and normoxia. Images were shown at magnification 400X. Numbers of tubes per file are shown. The data shown are the mean ± SD of independent experiments, n = 3. *P < 0.05; **P < 0.01 Student's 2-tailed t test or one-way ANOVA.

Fig 3: Histological confirmation of the increased expression of CXCL14 in IPF. Representative images of three IPF lung tissue sections (A–L) and three normal tissue sections (M–X) are stained for the presence of CXCR4 and CXCL14 with dapi counter stain. Increased expression of CXCL14 is observed in IPF only (C,G,K). The overlay denoted in yellow identifies regions of co-localization between CXCL14 and CXCR4 in regions of fibrosis (D,L). Wide spread CXCR4 is seen in all tissue (B,F,J,N,R,V) and specifically in the airways of both IPF (F) and normal tissue (V). CXCL14 is not seen in the airways of selected tissue (G,W).

Fig 4: Expression of tumor markers. a Expression of SDF1a levels after treatments and differences with tumor size. Analyses of linear regression performed in the sunitinib and bevacizumab treated tumors showed a significant correlation in these groups (r = 0.844, P = 0.0002). b Hemoglobin content in treated and control tumors analyzed according to tumor size. Analyses of linear regression performed in the sunitinib and bevacizumab treated tumors showed r = 0.69 and P = 0.016. c Western blotting determination of VEGF-A, HIF1a, and GCSF. d Growth curved derived from U87MG cell suspensions obtained from U87MG xenografts treated for 35 days with bevacizumab and scored as indicated in the text in high CXCR4 expression (cell suspensions from larger tumors) and low CXCR4 expression (cell suspensions from smaller tumors) and grown for 45 days with or without bevacizumab

Fig 5: 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.