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. 2022 Jul 20;298(9):102293. doi: 10.1016/j.jbc.2022.102293

Figure 3.

Figure 3

Roles of Tardbp on neurite extension in cultured neuronal cells.A, inhibition efficacy of shRNA expression constructs for Tardbp knockdown. ShRNA-control, Tardbp-shRNA1, Tardbp-shRNA2, or Tardbp-shRNA3 expression plasmids were cotransfected with plasmids expressing a FLAG-tagged Tardbp and a GFP into HEK293T cells. Western blot analysis was performed using anti-FLAG and anti-GFP antibodies. GFP was used as an internal transfection control. Tardbp-shRNA1, Tardbp-shRNA2, and Tardbp-shRNA3 suppressed Tardbp expression. B and C, ShRNA-control, Tardbp-shRNA1, Tardbp-shRNA2, or Tardbp-shRNA3 expression plasmids were cotransfected with a plasmid expressing FLAG-tagged EGFP into Neuro2a cells. Cells were immunostained with anti-FLAG and anti-α-tubulin antibodies. Nuclei were stained with DAPI (B). The length of the longest neurite of the transfected cells was measured (C). Data are presented as the mean ± SD. ∗p < 0.05, ∗∗p < 0.01 (one-way ANOVA followed by Tukey-Kramer test), n = 3 experiments. A total of 63, 25, 59, and 55 cells were measured in shRNA-control, Tardbp-shRNA1, Tardbp-shRNA2, and Tardbp-shRNA3, respectively.