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HEP_26230_sm_SuppFig1.tif1112KSupporting Information Figure 1. Accumulation of Hedgehog-responsive liver progenitors progressively declines as healthy murine livers mature. (A-F) Embryonic liver sections (embryonic day 13 [E13D] and 14 [E14D]) and liver sections from post-natal livers (post-natal week 3 [PN3W] and 12 [PN12W]) of healthy mice were stained for markers of liver progenitors (Sox9 and AFP) and the hedgehog-regulated transcription factor (Gli2). (A) Representative photomicrographs of Sox9-positive cells. (B) Quantitative analysis of Sox9-positive cells per high-powered field. (C) Representative photomicrographs of AFP-expressing cells. (D) Quantitative analysis of AFP-positive cells expressed as a percentage of stained cells per high-powered field. (E) Representative photomicrographs of Gli2-positive cells. (F) Quantitative analysis of Gli2-positive cells per high-powered field. For each marker, liver sections from 3 mice/time point were analyzed. On each section, positively-stained cells were counted in 20 randomly chosen high-powered (200x) fields to generate an average number of positively-stained cells/mouse; mean +/-SEM results were calculated by summing data from all mice sacrificed at each time point, analyzed using one-way ANOVA, and graphed; *p<0.05 as indicated.
HEP_26230_sm_SuppFig2.tif737KSupporting Information Figure 2. Accumulation of ?SMA-positive cells and Hedgehog ligand-producing cells progressively decline as healthy murine livers mature. (A-D) Embryonic liver sections (E13D, E14D) and liver sections from post-natal livers (PN3W, PN12W) of healthy mice were stained for αSMA (A-B) or Indian Hedgehog ligand (IHh, C-D). (A) Representative photomicrographs of αSMA-positive cells. (B) Quantitative analysis of αSMA-positive cells expressed as a percentage of stained cells per high-powered field. (C) Representative photomicrographs of IHh-expressing cells. (D) Quantitative analysis of IHh-positive cells expressed as a percentage of stained cells per high-powered field. For each marker, liver sections from 3 mice/time point were analyzed. On each section, positively-stained cells were counted in 20 randomly-chosen high-powered (200x) fields to generate an average number of positively-stained cells/mouse; mean +/-SEM results were calculated by summing data from all mice sacrificed at each time point, analyzed using one-way ANOVA, and graphed; *p<0.05 as indicated.

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