Fig. 2
Loss of Smarca5 in vivo during normal cerebellar development and SHH-MB formation. (a) H&E staining and immunohistochemistry for SMARCA5 in the cerebellum of Math1-cre (left panel), Math1-cre::Smarca5Fl/+ (middle) and Math1-cre::Smarca5Fl/Fl (right panel) mice at P5. (b) Immunohistochemistry for Ki67 and Cleaved caspase 3 (CC3) in the EGL of cerebella from mice with indicated genotypes at P5. Black arrowheads indicate cleaved Caspase-3 positive cells. (c) Quantification of Ki67 and cleaved Caspase 3-positive cells in the EGL of mice shown in (b) (n = 4, Fisher´s exact test). (d) Representative H&E stainings of whole cerebella from Math1-cre (left panel), Math1-cre::Smarca5Fl/+ (middle panel) and Math1-cre::Smarca5Fl/Fl (right panel) mice at P21. (e) Representative H&E stainings of cerebellar tumors and immunohistochemistry for SMARCA5, Ki67 and cleaved Caspase 3 in tumors from mice with indicated genotypes at P5. (f) Quantification of Ki67 (left) and cleaved Caspase 3 (right) positive cells in tumors from mice shown in (e) (n = 3, Fisher´s exact test). (g) Kaplan-Meier curves showing survival of Math1-cre::SmoM2Fl/+ (n = 11), Math1-cre::Smarca5Fl/+::SmoM2Fl/+ (n = 11) and Math1-cre::Smarca5Fl/Fl::SmoM2Fl/+ (n = 11) mice. Significance in survival is indicated using log rank (Mantel-Cox) test. (h) H&E-stained cerebellar tumors from Math1-cre::SmoM2Fl/+ (left) and Math1-cre::Smarca5Fl/Fl::SmoM2Fl/+ (right) mice at different endpoints. Immunohistochemistry for SMARCA5 (Sm5) (upper panel) and Ki67 (lower panel) in tumors from these mice are indicated. (i) Fraction of SMARCA5-negative cells in cerebellar tumors from Math1-cre::Smarca5Fl/Fl::SmoM2Fl/+ mice from P5 to P27. 4x magnification, scale bar, 500 μm; 20x magnification, scale bar, 50 μm; 40x magnification, scale bar, 20 μm. EGL, external granular layer; ML, molecular layer; IGL, internal granular layer. All graphs display mean ± SD. *p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.
