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. Author manuscript; available in PMC: 2015 Jul 23.
Published in final edited form as: Methods Mol Biol. 2014;1213:229–243. doi: 10.1007/978-1-4939-1453-1_19

Fig. 2.

Fig. 2

Gating strategy for FACS purification of adipogenic progenitor cells from a heterogeneous muscle cell population. (a) In the FSC vs. SSC plot, draw a polygonal region (R1) around the events that represent intact cells, thereby excluding debris. (b) Perform duplet discrimination by gating for single cells (R2) in FSC-H vs. FSC-A plot as indicated. (c) Calcein and Propidium iodide (PI) are used to discriminate between live (Calcein+/PI−, R3) and dead cells (Calcein−/PI+). (d) Exclude hematopoietic cells and endothelial cells by selecting cells that are CD11b−/CD45−/CD31− (R4: lineage negative (Lin−) cells) in the PE/Cy7 vs. FITC plot. (e) Select MusAPCs marked as the Sca1+/Lin− population (R6) for sorting. For optional analysis/collection of myogenic progenitors, define a gate for the Sca1−/Lin− population (R5). (f) To enrich satellite cells within the myogenic Sca1−/Lin− population, select the Integrin ß1 positive population (R7) and apply a specific selection marker for satellite cells (see Note 8)