Anti tubulin β3 primary antibody tuj1

Two days after in vivo electroporation of fluorescent siRNA control, mice were perfused and the electroporated L4/5 DRGs were collected. Frozen DRG sections of 10 μ were obtained with a cryostat and warmed on a slide warmer at 37C for 1 hr. After being blocked with PBS containing 10% goat serum and 0.3% Triton X-100 at room temperature for 1 hr, the sections were immunostained with anti-tubulin β3 primary antibody (TUJ1, 1:500, Biolegend) overnight at 4C, followed by Alexa Fluor 488 conjugated secondary antibody (1:500, Thermo Fisher Scientific) at room temperature for 1 hr. All antibodies were diluted with the blocking buffer. Four times of 15-min wash with PBS containing 0.3% Triton X-100 was performed following each antibody incubation. The DRG sections were mounted with Fluoroshield histology mounting medium (Sigma-Aldrich) and imaged with the inverted fluorescent microscope mentioned above. Four or five non-adjacent sections from each DRG were used for analysis. For each DRG, the transfection rate was calculated by dividing the of number of red fluorescent siRNA control⁺/TUJ1⁺ cells by the number of TUJ1⁺ cells.