Extended Data Fig. 6: Generation and phenotypes of single-Stag GSCs.
From: The mitotic STAG3–cohesin complex shapes male germline nucleome
a, (Top) Scheme for PAM and guide RNA sequences in the Stag1 exon4 (left) or in the Stag2 exon8 (right). (Bottom) Sequences of the targeted loci in the Stag1/2 double-knockout (dKO) (Stag3-only) and Stag1/3 dKO (Stag2-only) lines (left) or in the Stag1/2 dKO (Stag3-only) and Stag2/3 dKO (Stag1-only) lines (right). b, The protein abundance of STAG1/2/3 was measured by in-gel digestion followed by mass spectrometry (MS) in the indicated cell lines. The protein abundance was calculated from the peak area intensities of the top 3 unique peptides that only map to the protein and do not map to other identified proteins. The protein abundance was represented using iBAQ. Two technical replicates (dots) were analyzed and the averages are shown as bars. c, Doubling time (hour, averages as bars) of WT (n = 4) and single-Stag GSCs (each n = 2). While all single-Stag GSCs appeared to show slower propagation compared to WT GSCs, we did not perform statistical analysis, since independent each single-Stag GSC clone numbers we analyzed were two. d, Representative IF images of GFP (green), SYCP3 (red) and DAPI (grey) in a testis transplanted with Stag3 KO GSCs. Bar, 50 μm. Two slides obtained from two independent mice, were analyzed. e, Representative IF images of GFP (green), SYCP3 (red) and DAPI (grey) in a testis transplanted with Stag3-only GSCs. Bar, 50 μm. Five slides obtained from two independent mice, were analyzed.
