Figure 7.

Assessment of the astroglial differentiation of NPCs by calculation of the GFAP-immunointensity (GFAP-i; pixel intensity (pi)/mm²) per GFP⁺ cells (GFAP-i/NPC index). (A) Immunofluorescence staining for GFAP (blue) indicating glial differentiation, GFP⁺, and GFAP⁺/GFP⁺ cells. (B) Under normal growing conditions, but also under inflammatory stress, Cyclopamine-treatment and thus blockage of the Shh-pathway results in a significant increase of the GFAP-i/NPC index, and therefore, significantly more astrocytes compared to untreated cells (Cyclo only vs. control; p = 0.0009 and Cyclo+LPS vs. LPS only; p = 0.004, respectively) and Shh-treated cells (Cyclo only vs. Shh only; p = 0.002 and Cyclo+LPS vs. Shh+LPS; p = 0.038, respectively). Experiments performed in triplicate (n = 3), data presented as mean ± SEM, one-way-ANOVAs with post hoc Tukey-HSD tests performed for statistical analysis (* p < 0.05, ** p < 0.01, *** p < 0.001).