Journal: Regenerative Therapy

Article Title: Enhanced axon outgrowth of spinal motor neurons in co-culturing with dorsal root ganglions antagonizes the growth inhibitory environment

doi: 10.1016/j.reth.2023.11.013

Figure Lengend Snippet: A: The SMNs in dispersed culture on the 5th day (upper). 3D printed mold and PDMS template for SMNA preparation (lower left) and the RFP-transduced SMNA plated on the 3rd day (lower right). B: The cultured SMNs were immunofluorescently identified as positive for ChAT, Tuj1 and NeuN. C: Different degrees of axonal connections shown at two populations of SMNAs plated at various distances. Robust axonal connections formed in 2 mm of distance (upper), a small number of axonal connections formed in 3 mm of distance (middle) and no axonal connection was observed in 4 mm of distance (lower). Scale Bar: A, B: 100 μm C: 200 μm.

Article Snippet: Then, the rabbit anti-ChAT antibody (Beyotime, AF6497, 1:500), mouse anti-neuron-specific Class III β-tubulin antibody (Tuj1, Covance, MMS-435P, 1:500), or rabbit anti-NeuN (Proteintech, 26975-1-AP, 1:500) was added to the cell slides for incubation overnight at 4 °C, followed by incubation for 30 min at 37 °C with Cy3-labelled goat anti-rabbit IgG (H & L) (Beyotime, A0516, 1:500), Cy3-labelled goat anti-mouse IgG (H & L) (Beyotime, A0521, 1:500) or Dylight 488-labelled Goat anti-rabbit IgG (H & L) (Abcam, ab96883, 1:1000) as the second antibody, respectively.

Techniques: Cell Culture