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Files in this Data Supplement:
Fig. S1. Gross morphological changes in the cortex of E12.5 Lifr-/- embryos relative to control littermates are suggestive of defects related to growth. Measurements of the area of the cortex revealed a 17% decrease in size in Lifr-/- embryos (A,B; paired t-test **P=0.01; n=5). A concurrent decrease in the thickness of the cortical plate was observed, as shown by b-tubulin III staining (C,D). The number of dying cells in the LGE of Lifr-/- embryos indicated by TUNEL labeling was normalized to the area of the LGE (F; 0.3±0.03 cells/unit area) and no difference was observed relative to Lifr+/+/Lifr+/- embryos (E; 0.3±0.07 cells/unit area; n=3; arrows indicate positive cells, asterisks indicate non-specific autoflorescence). The number of BrdU+ cortical precursors in Lifr+/+/Lifr+/- embryos (C; 1502±96) was significantly higher than Lifr-/- embryos (D; 1029±21; paired t-test **P=0.009; n=3). The number of surface pHH3+ cells (VZ precursors) was decreased by 28% in Lifr-/- embryos relative to Lifr+/+/Lifr+/ littermates (I,J; paired t-test; *P=0.009; n=5); however, the number of non-surface pHH3+ cells (SVZ precursors) was not significantly different. Scale bar: in A, 100 mm for A,B; C, 100 mm for C,D; in E, 100 mm for E-H; in I, 50 mm for I,J.
Fig. S2. The number of dividing VZ precursors in the E12.5 Lifr-/- spinal cord is not different from control littermates. Analysis of the upper thoracic region of the spinal cord in E12.5 Lifr-/- embryos (B) revealed no gross morphological differences relative to controls (A; n=4) and the number of BrdU-positive cells in the Lifr-/- (H; 436±87) embryonic spinal cord VZ was not different from controls (G; 468±126; n=3). Scale bar: 50 mm.
Fig. S3. The activation of CNTF/Lif/gp130 signaling decreased the number of proliferating precursors in the VZ of E14.5 spinal cord explants. CNTF treatment (B; 8.4±0.8) decreased the number of pHH3 cells in explant cultures of the upper thoracic E14.5 spinal cord by 51% relative to basal media treatment (A; 17.1±1.9; paired t-test **P=0.0001; n=4). Scale bar: 100 mm.
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