Figure 3. LIF/STAT3 activation increases the formation of bRG cells and stimulates astrogliogenesis.

(A-C) Cerebral organoids immunostained for general RG (PAX6/SOX2), bRG (HOPX), IP (TBR2), and dividing RG (pVIM) cell markers. (D) Thickness of the SVZ in μm and number of bRG. bRG were counted as abventricular PAX6⁺ and pVIM⁺ cells. Data are represented as mean ± SEM. (E) Antibody staining for GFAP+/HEPACAM⁺ astrocytes or CTIP2⁺ excitatory neurons in control versus LIF-treated organoids. (F) Cerebral organoids and human fetal cortex immunostained for CTIP2⁺ lower layer neurons, BRN2⁺ upper layer neurons, and LAMININ⁺ basement membrane production. Arrowheads denote blood vessels. (G) W22 organoids with or without LIF treatment immunostained for the cell death marker clCAS3, CTIP2⁺ or TBR1⁺ lower layer neuronal markers, and SATB2⁺ or CUX1⁺ upper layer neuronal markers. (H) Relative cortical plate position of CTIP2+, TBR1+, SATB2+, or CUX1⁺ neurons in W22 organoids with or without LIF. Values represent median ± 95%CI. Total number of neurons counted from 3 independent experiments, Control: CTIP2⁺ n=,837 TBR1⁺ n=1088, SATB2⁺ n=836, CUX1⁺ n=718; LIF: CTIP2⁺ n=931, TBR1⁺ n=1038, SATB2⁺ n=765, CUX1⁺ n=546; n.s. no significance, ****p < 0.0001, Statistical analysis compares indicated neuronal markers to the CTIP2⁺ group, Kruskal-Wallis test with Dunn's Correction. All scale bars: 50 μm. See also Figures S4 and S5.