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. 2013 Mar 15;123(4):1833–1843. doi: 10.1172/JCI65822

Figure 4. SALL-4–positive human spermatogonia do not express HLA-ABC or CD49e.

Figure 4

(A) To determine whether human spermatogonia express HLA-ABC, human testicular cell suspensions were stained with APC-conjugated HLA-ABC antibodies and sorted into positive and negative fractions by FACS. Negative gates were defined by analysis of human testis cells using APC-conjugated isotype control antibodies. (B–E) Following FACS, each fraction of cells was fixed and immunocytochemistry was performed to assess SALL-4 expression; then, fractions were counterstained with DAPI to quantify total cells. (B) The percentage of cells in each unsorted and sorted fraction that displayed SALL-4 staining (SALL-4+ green cells/DAPI-stained total cells). (F–J) A similar experiment was conducted using APC-conjugated CD49e antibodies. Scale bar: 50 μm (C–E and H–J). Bars in B and G indicate the mean percentage of SALL-4–positive cells (SALL-4–positive cells/total cells) in each fraction. Error bars in B and G represent SEM from 3 replicate sorting experiments. *P < 0.001, compared with unsorted cells.