Figure 6

hCDNF inhibits ER stress markers in astroglia in the acute MPTP model. (A,B) Z-stack images of immunofluorescence double staining for GFAP (red), GRP78 (cyan), DAPI (blue) and GFP or hCDNF (green) in the STR (upper panel, A) and SN (lower panel, B). The dotted white rectangles indicate the enlarged areas. Scale bar: 50 µm for the z-stack images and 25 µm for the enlarged images. (C) Quantification of the GFAP + GRP78 pixel area in the STR and SN. (D,E) Western blot analysis of GRP78 and β-actin expression in the STR and SN. (D) Quantification of GRP78 expression in the STR and normalization to β-actin expression. (E) Quantification of GRP78 expression in the SN normalized to β-actin expression. (F,G) Z-stack images of immunofluorescence double staining for GFAP (red), CHOP (cyan), DAPI (blue) and GFP or hCDNF (green) in the STR (upper panel, F) and SN (lower panel, G). The dotted white rectangles indicate the enlarged areas. Scale bar: 50 µm for the z-stack images and 25 µm for the enlarged images. (H) Quantification of the GFAP + CHOP pixel area in the STR and SN. (I,J) Western blot analysis of CHOP and β-actin expression in the STR and SN. (I) Quantification of CHOP expression in the STR normalized to β-actin expression. (J) Quantification of CHOP expression in the SN normalized to β-actin expression. (C) STR: n = 15 images/4 mice for the PBS group, n = 13 images/4 mice for the PBS + MPTP group, n = 15 images/4 mice for the AAV-GFP + MPTP group, and n = 19 images/4 mice for the AAV-hCDNF + MPTP group; SN: n = 12 images/4 mice for the PBS group, n = 12 images/4 mice for the PBS + MPTP group, n = 14 images/4 mice for the AAV-GFP + MPTP group, and n = 14 images/4 mice for the AAV-hCDNF + MPTP group. (H) STR: n = 13 images/4 mice for the PBS group, n = 14 images/4 mice for the PBS + MPTP group, n = 12 images/4 mice for the AAV-GFP + MPTP group, and n = 19 images/4 mice for the AAV-hCDNF + MPTP group; SN: n = 13 images/4 mice for the PBS group, n = 12 images/4 mice for the PBS + MPTP group, n = 12 images/4 mice for the AAV-GFP + MPTP group, and n = 12 images/4 mice for the AAV-hCDNF + MPTP group. (D,E,I,J) n = 6 for the PBS group, n = 6 for the MPTP group, n = 5 for the AAV-GFP + MPTP group, and n = 6 for the AAV-hCDNF + MPTP group. (C–E,G,I,J) One-way ANOVA followed by Tukey’s multiple comparisons post hoc test. All the data are expressed as the mean ± S.E.M. *p < 0.05, **p < 0.01, and ***p < 0.001.