Fig. 4

hMSC treatment preserves the protein levels of follistatin-like 1 (FSTL1) and transforming growth factor beta 1 (TGF-β1) in the cerebellum of SCA2 mice. A, B Western blot analysis of FSTL1 and TGF-β1 in cerebellum tissues obtained from 10-, 25-, and 50-week-old WT and SCA2 mice. **p < 0.001 and ***p < 0.001 vs. 10-week-old WT mice (two-way ANOVA with Tukey’s post hoc analysis; n = 6 for each group). Full-length blots are presented in supplementary Fig. 5; Fig. 4A. C Double immunofluorescence staining was performed for calbindin and FSTL1 as well as calbindin and TGF-β1 in the cerebellum of 50-week-old WT and SCA2 mice. The scale bar represents 20 μm. The dashed line differentiates between the granule and Purkinje cell layers. D The fluorescence intensity for the quantitative co-localization of calbindin with FSTL1 and TGF-β1 was measured. ***p < 0.001 vs. untreated WT mice (t-test with Tukey’s post-hoc analysis). E Double immunofluorescence staining was performed for NeuN and FSTL1 as well as NeuN and TGF-β1 in the cerebellum of 50-week-old WT and SCA2 mice. The scale bar represents 20 μm. The dashed line differentiates between the granule and Purkinje cell layers. F The fluorescence intensity for the quantitative co-localization of NeuN with FSTL1 and TGF-β1 was measured. ***p < 0.001 vs. untreated WT mice (t-test with Tukey’s post-hoc analysis). G, H Western blot analysis of FSTL1 and TGF-β1 in the cerebellum after 24 weeks of hMSC administration (50-week-old mice). ***p < 0.001 vs. untreated WT mice; ^###p < 0.001 vs. untreated SCA2 mice (two-way ANOVA with Tukey’s post hoc analysis; n = 6 for each group). Full-length blots are presented in supplymentary Fig. 5: Fig. 4G. I Schematic representation of the effects of hMSCs in the SCA2 model. hMSC administration upregulates the expression of FSTL1, which in turn reduces inflammation and is accompanied by behavioral and physiological improvements in the SCA2 phenotype. The figure was created using BioRender.com (Agreement number: BG25IEMZTA)