Supplementary Figure 6: Biochemical and cellular characterization of the Mmp24-KO mice.

(a) Upper panel: relative mRNA levels of Cdh2 normalized to the ribosomal protein L32 in semiquantitative RT-PCRs in total RNA extracts of primary neurospheres obtained from Mmp24-KO and wild-type animals and grown for 3 days. Mean ± s.e.m. Two tailed unpaired Student’s t Test. Dots represent extracts from cultures established from independent mice. Statistics source data can be found in Supplementary Table 2. Bottom panel: representative image of the bands quantified. (b) Upper panel: densitometric quantification of β-catenin in total SEZ protein extracts from Mmp24-KO and wild type controls normalized to a loading control. Mean ± s.e.m. Two tailed paired Student’s t Test. Dots represent extracts from cultures established from independent mice. Statistics source data can be found in Supplementary Table 2. Bottom panel: representative immunoblots for β-catenin in total protein extracts of primary neurospheres obtained from Mmp24-KO and wild-type animals and grown for 3 days. β-actin is shown for loading control. (c) Representative immunoblot for N-cadherin, the intracellular active domain of Notch (NICD), and β-actin for loading control in extracts from Mmp24-KO neurospheres lentivirally infected for the expression of FLAG-tagged MT5-MMP or with the empty vector (GFP). (d) FLAG immunoblot showing expression of MT5-MMP in infected cells. See Supplementary Fig. 8 for uncropped western blots. (e) Relative number of primary neurospheres obtained from the SEZ of wild-type and mice deficient in MT5-MMP activity. Mean ± s.e.m. Two tailed unpaired Student’s t Test. Dots represent neurosphere cultures obtained from independent mice. Statistics source data can be found in Supplementary Table 2. (f) Percentage of S100β⁺ cells within the CldU-LRC⁺ population in the SEZ of wild-type and Mmp24-KO mice, injected with the nucleotide every 2 h during a 12 h period, and allowed to survive for 28 days before euthanasia. Mean ± s.e.m. Two tailed unpaired Student’s t Test. Each dot represents a mouse. Statistics source data can be found in Supplementary Table 2. (g) Representative images of the immunodetection of S100β positive (arrowheads) and CldU-LRC double positive cells (arrows). Scale bars: 10 μmm.