Figure 3. In a mixed population of MSCs the αSMA-RFP- cells contain the stem cells and RFP+ cells express a typical MF marker.
(A) FACS for αSMA-RFP+ cells in freshly isolated whole BM of 6, 9, 12, 15 and 18 month old αSMA-RFP+ mice and αSMA-RFP+ mice with H. felis infection (red) and 6, 12 and 15 month old IL-1β/αSMA-RFP+ transgenic mice (green) and 6, 9 and 12 month old αSMA-RFP+ mice with H. felis infection and MNU treatment (purple).
(B) αSMA and RFP staining of the BM of C57/B6 (lower row) or αSMA-RFP (upper row) mice before (control) and after 14 months of H. felis infection. Arrows indicate cells staining positive. (*=p< 0,05, data are represented as mean +/− SEM) See also Figure S3 and Table S2
(C) CFUs from 1000 plated RFP+ (red), RFP- (green) or RFP+/− (black) cells were counted after 14 days culture.
(D) Adipocyte (brown) and osteoblast (orange) differentiation of RFP+, RFP- or RFP+/− cells was measured after 14 days culture in differentiation mediua. After Oil Red-O or Alizarin Red staining % of differentiated cells was calculated.
(E) Representative pictures of RFP+/− cells after differentiation experiments (left as control) and after adipocyte (middle, with Oil Red-O staining) or osteoblast (right with Alizarin Red staining) differentiation.
(F) Quantitative expression of αSMA or collagen in RFP+ (red), RFP- (green) or RFP+/− (black) cells at passage 5 after a single RFP sorting.
(G) Representative pictures of αSMA, FSP1, collagen, and vimentin staining in RFP+, RFP− or RFP+/− cells. (*=p< 0.05, **=p<0.01 data are represented as mean +/− SEM) (See also Figure S3)