Fig. 3: In situ characterization of hybrid cells. | Nature Communications

Fig. 3: In situ characterization of hybrid cells.

From: Collagen signaling and matrix stiffness regulate multipotency in glandular epithelial stem cells in mice

Fig. 3

a Confocal imaging of immunostaining of MG organoid from K5CreER/td-Tomato mice in Matrigel, Soft, Col1, and Stiff gel 1 week after TAM administration using anti-K14 (White) and anti-K8 (Green) antibodies. TOM in red, Hoechst in blue. Arrows indicate hybrid cells (TOM + K14 + K8+) and fully differentiated LCs (TOM + K14− K8+). Scale bars, 10 μm. FACS quantification of TOM+ hybrid (K5 + K8+) on the total cells (b) and TOM+ BCs on the total BCs (c) in MG organoids at different conditions indicated 1 week after TAM administration. n = 7 independent experiments. d Representative images of immunostaining of the organoid derived from K5CreER/td-Tomato mice in Matrigel, Col1, Soft, and Stiff gel 2 days after TAM administration. TOM in red, K8 in green, K14 in white, Hoechst in blue. Arrows indicate hybrid cells (TOM + K14 + K8+). Scale bars, 50 μm. e Representative FACS plot of K5 and K8 expression on TOM+ cells from MG organoids derived from K5CreER/td-Tomato 2 days after TAM administration. MG organoids are embedded in the different gels indicated. BC: K5-high K8-low; LC: K5-low K8-high; Hybrid: K5 + K8+. f Quantification of TOM+ LCs on total LCs, Tom+ Hybrid on total cells, Tom+ BCs on total BCs in MG organoids at different conditions indicated 2 days after TAM administration. n = 3 independent experiments. For (b, c, and f), graphs are mean ± s.e.m. p-values are derived from two-sided unpaired t-test. Source data are provided as a Source Data file.

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