Cd117 c kit

Immunofluorescence staining was performed as described [33 (link), 83 (link), 84 (link)]. Briefly, exponentially growing cells were fixed with 4% paraformaldehyde (PFA) for 10 min, washed with PBS and permeabilized with 1% NP-40 for 10min at room temperature. After being blocked with 10% donkey serum (Jackson Immuno-Research Laboratories, West Grove, PA) for 1h at room temperature, cells were incubated with various primary antibodies, including CD29, CD73, BMPRII, CD90, CD117/c-kit, CD105/endoglin, or BMPR-II antibody (all from Santa Cruz Biotechnology) for 1h at room temperature. Cells were washed with PBS and incubated with FITC-labeled secondary antibodies (Jackson ImmunoResearch Laboratories) for 30 min. DAPI (Invitrogen) was used to visualize nuclei. Stains were examined under a fluorescence microscope. Negative control cells were performed under the same conditions without primary antibodies. Representative images from at least three independent staining experiments are shown.

Immunofluorescence staining was performed 43, 51. Briefly, cells were seeded in 24‐well plates overnight, fixed with 4% paraformaldehyde, permeabilized with 1% NP‐40 and blocked with 10% donkey serum (Jackson Immuno‐Research Laboratories, West Grove, PA, USA), followed by incubating with CD73, CD105/endoglin, CD90/Thy‐1, CD166/ALCAM, BMPR‐II, CD117/c‐kit or CD29/Integrin β1 antibody (Santa Cruz Biotechnology, Dallas, TX, USA) for 1 hr at room temperature, as previously reported 43, 56. After being washed, cells were incubated with Texas Red or FITC labelled secondary antibody (Jackson ImmunoResearch Laboratories) for 30 min. Cell nuclei were counterstained with DAPI. Stains without primary antibodies were used as negative controls. Fluorescence images were recorded under an inverted fluorescence microscope.

Immunofluorescence staining was performed as described [22] (link), [24] (link), [35] (link), [37] (link), [40] (link), [52] . Briefly, cells were fixed with methanol, permeabilized with 1% NP-40, and blocked with 10% BSA, followed by incubating with CD73, CD44, CD90, CD117/c-kit, CD29, CD133, CD105/endoglin, CD166/ALCAM, or BMPR-II antibody (Santa Cruz Biotechnology) for 1 hr at room temperature. After being washed, cells were incubated with Texas Red-labeled secondary antibody (Santa Cruz Biotechnology) for 30 min. Cell nuclei were stained with DAPI. Stains were examined under a fluorescence microscope. Stains without primary antibodies, or with control IgG, were used as negative controls.