Fig. 9. RBM5 Overexpression Increases Vulnerability of Primary CNS Cells to Stretch-Injury.

(a) Total protein stain shows equal protein loading/transfer across groups of non-transduced DIV9 cortical neurons subjected to a single or multiple stretch-injuries. Western blots show levels of RBM5 and RBM10 before and after injury. (b) DIV0 cortical neurons were transduced with 30MOI lentivirus to deliver either an empty-vector (control) or RBM5 open reading frame (ORF, n=4/group). Both virus-control and RBM5-ORF groups were subjeted to a single 54% biaxial stretch on DIV9, and cell culture media plus protein extracts were harvested 24h later (n=4/group). Western blots shows RBM5, anti-FLAG, and SBDPs levels. (c) Densitometric analysis of SBDP levels in injured 30MOI control vs. RBM5-ORF groups (n=4/group). (d) 24h post-injury LDH levels in 30MOI control vs. RBM5-ORF groups (n=4/group). (e) Western blots show RBM10, NFL, and Tau in injured 30MOI control vs. RBM5-ORF groups (n=2/group). (f) DIV0 cortical neurons were transduced with 60MOI lentivirus to deliver either an empty-vector (control) or RBM5-ORF (n=4/group). Both virus-control and RBM5-ORF groups were subjeted to a single 54% biaxial stretch on DIV9, and cell culture media plus protein extracts were harvested 24h later (n=4/group). Western blots shows RBM5 and SBDPs levels. (g) Densitometric analysis of SBDP levels in injured 60MOI control vs. RBM5-ORF groups (n=4/group). (h) 24h post-injury LDH levels in 60MOI control vs. RBM5-ORF groups (n=4/group). Data were anlayzed by unpaired T-test. Data were significant at p<.05. Graphs show mean +SEM. (*) = p<.05; (***) = p>.0001.