Figure 3
Heat-labile soluble factor(s) mediates YAP 5SA-induced astrogenesis in vitro. (A) Immunostaining using anti-GFAP after in vitro differentiation of co-cultured cells. E13.5 neural progenitor cells transduced with YAP 5SA retroviruses were mixed with untransduced neural progenitor cells at a ratio of 1:5 (transduced:untransduced) and then cultured in differentiation medium for 3 days. Quantification of (A) is shown in (B). (C) GFP (green) and GFAP (red) double immunostaining of cells differentiated under the same experimental conditions as (A). (D) Untransduced E13.5 neural progenitor cells were cultured in differentiation medium prepared by mixing conditioned medium (CM) of YAP 5SA-transduced neural progenitor culture and fresh differentiation medium in a 1:1 ratio. CMHI, heat-inactivated (56 °C for 30 min) CM. (E) Quantification of (D). The DAPI nuclear counterstain is shown in blue in (A, D). Scale bars, 100 μm for (A, D), and 200 μm for (C). Student’s t-test was used to determine statistical significance. **P < 0.01, ***P < 0.001; n ≥ 3.
