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. 2023 Aug 23;12:RP85380. doi: 10.7554/eLife.85380

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© 2023, Wang, Jin et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 2. — (A) Schematic illustration of the lineage tracing workflow for FOXC2⁺ cells. (B) Immunostainings for DAPI (blue) and FOXC2 (red) at day 3 post TAM induction. Scale bar, 50 μm; d, day. (C) Fluorescence-activated cell sorting (FACS) analysis of GFP⁺ populations derived from Rosa26^{LSL-T/G/LSL-T/G} or Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice at day 3 post TAM induction. (D, E) The recipient mice testes (D) and colony numbers (E) 2 months after transplantation (n=10) of the FACS-sorted GFP⁺ cells from the Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice 3 days after TAM diet and the FACS-sorted THY1⁺ cells from adult mice. Scale bar, 1 mm; values, mean ± s.e.m.; p-values were obtained using two-tailed t-tests (****p-value <0.0001). (F) Immunostaining for DAPI (blue), ZBTB16/FOXC2 (red), and GFP (green) at week 1 post TAM induction (scale bar, 50 μm). (G) Seminiferous tubules of Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice 2, 4, and 6 weeks post TAM induction. Scale bar, 50 μm. (H) Testes (scale bar, 1 mm), seminiferous tubules, and epididymis (scale bar, 50 μm) at months 4, 7, and 12 post TAM induction in Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice. (I, J) The GFP⁺ patches (I) and progeny (J) population dynamics (n=10). Values, mean ± s.e.m. (K) Immunostainings for DAPI (blue), EOMES (red), GFRA1 (red), or PAX7 (red) in GFP⁺ population at week 2 post TAM induction. Scale bar, 50 μm.

Figure 2—figure supplement 1. — (A) Schematic illustration of the lineage tracing workflow for FOXC2⁺ cells. (B) Immunostainings for DAPI (blue) and FOXC2 (red) at day 3 post TAM induction. Scale bar, 50 μm; d, day. (C) Fluorescence-activated cell sorting (FACS) analysis of GFP⁺ populations derived from Rosa26^{LSL-T/G/LSL-T/G} or Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice at day 3 post TAM induction. (D, E) The recipient mice testes (D) and colony numbers (E) 2 months after transplantation (n=10) of the FACS-sorted GFP⁺ cells from the Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice 3 days after TAM diet and the FACS-sorted THY1⁺ cells from adult mice. Scale bar, 1 mm; values, mean ± s.e.m.; p-values were obtained using two-tailed t-tests (****p-value <0.0001). (F) Immunostaining for DAPI (blue), ZBTB16/FOXC2 (red), and GFP (green) at week 1 post TAM induction (scale bar, 50 μm). (G) Seminiferous tubules of Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice 2, 4, and 6 weeks post TAM induction. Scale bar, 50 μm. (H) Testes (scale bar, 1 mm), seminiferous tubules, and epididymis (scale bar, 50 μm) at months 4, 7, and 12 post TAM induction in Foxc2^iCreERT2/+;Rosa26^{LSL-T/G/LSL-T/G} mice. (I, J) The GFP⁺ patches (I) and progeny (J) population dynamics (n=10). Values, mean ± s.e.m. (K) Immunostainings for DAPI (blue), EOMES (red), GFRA1 (red), or PAX7 (red) in GFP⁺ population at week 2 post TAM induction. Scale bar, 50 μm.