Fig 4.
CRISPRa-engineered ASC sheets stimulated neuron proliferation and neurite outgrowth. (A-F) DRG neuron proliferation and extension as detected by immunolabeling specific for NeuN. (G) Neuron numbers. (H) Average neurite length. DRG neurons were seeded onto cover glasses (5×105 cells/glass) for 2 days and a wound gap was created using a cell scrapper. The neurons on the cover glass were transferred to a new 6-well plate and cultured alone as a negative control (NC group) or co-cultured with the mock-transduced (Mock group) or Bac-Cre/Bac-LECW-transduced (MOI=100/150, CRISPRa group) rat ASC sheets. After 5 days, the neurons were subjected to NeuN-specific immunostaining. Four fields in each cover glass were randomly chosen and the numbers of neurons extending through the gap were counted using ImageJ software. The average neurite lengths were also calculated using ImageJ software. The data represent means±SD of 3 independent culture experiments.