Fig 1: Deletion of Wls from Col1a1-expressing cells does not affect Wls expression or the secretion and senescence of HSCs in the spleens of TBI-exposed mice. Four-week-old Wlsfl/fl and Col-Cre;Wlsfl/fl mice were exposed to sub-lethal TBI, and (A) splenic Wls mRNA and (B) Wnt3a and Wnt5a protein levels in spleen supernatants were determined 4 weeks post-TBI by qRT-PCR and ELISA, respectively (n = 6). At the same time post-TBI, (C) the number of HSCs and (D) MitoSox- or (E) C12FDG-positive HSCs in the spleens of those mice were determined by flow cytometry (n = 5). Col1a1-Cre activity was determined in (F) the trabecular and cortical zones of the femur and (G) the spleens of mutant and control mice 4 weeks post-TBI. The representative images in panel (F) show regions stained with X-gal (blue), which indicate the active Cre recombinase sites in the sections. Here, the images exhibiting the X-gal-specific intensity at average level among five different samples were represented. The p values in panels A and B were determined by unpaired Student’s t-test. The p values in panels C-E were calculated using unpaired non-parametric Wilcoxon t-test. ns, not significant.
Fig 2: TBI acutely reduces the numbers of peripheral WBCs and lymphocytes but not the BM levels of Wnt ligands, and that is ameliorated by osteoblastic Wls depletion. The levels of circulating (A) WBCs, (B) lymphocytes, (C) granulocytes, (D) RBCs, and (E) platelets in the mouse groups were measured using an automated complete blood cell counter at the indicated times (h) after TBI (n = 5). (F) The BM levels of Wnt3a and Wnt5a in Wlsfl/fl and Col-Cre;Wlsfl/fl mice were evaluated by IHC when they were 4 weeks of age. Representative data from five different samples are shown. Scale bars = 100 µm. (G) The area (%) positively stained with Wnt3a or Wnt5a in the IHC assay was calculated (n = 5). (H) Levels of Wnt3a and Wnt5a mRNA in whole BM lysate were determined by qRT-PCR (n = 5). Protein levels of (I) Wnt3a and (J) Wnt5a in the whole BM lysate were determined by ELISA at the indicated times after TBI (n = 5). The p values in all panels were determined by non-parametric Wilcoxon t-test. The superscriptsa-c indicate significant differences among the groups compared with the value of non-TBI control or mutant group by ANOVA. The symbol ‘*’ in panels I and J indicate significant difference at p < 0.043 by the Wilcoxon t-test. ns, not significant.
Supplier Page from Aviva Systems Biology for WNT3A ELISA Kit (Mouse) (OKEH03470)
Specificity: Natural and recombinant Mouse Protein Wnt-3a