Figure 5
From: TDP-43 knockdown impairs neurite outgrowth dependent on its target histone deacetylase 6
Rescue of neurite outgrowth in shHDAC6 cells by HDAC6 transfection. Parental SH-SY5Y cells (Ctrl) or stably transduced cells with shRNA against HDAC6 (shHDAC6, 1.5 × 105 TU) were transiently transfected with Myc-HDAC6 or control vector. A, Cells were lysed, electrophoresed and Western blots sequentially probed with antibodies against HDAC6 (top panel) and TDP-43 (middle panel). Anti-GAPDH probing (bottom panel) confirmed equal loading. B, cells were exposed to RA. After 1d, cells were transfected for 4h with Myc-HDAC6 or control vector and treated for another 3d with RA. B-E, Cells were fixed and labeled with Alexa568-phalloidin (red) and anti-Myc (green). Nuclei were counterstained with Hoechst (blue). B, shown are representative images. Scale bar corresponds to 20 μm. Quantifications of transfected, Myc-positive cells were performed for the parameters C, number of neurites per cell, D, number of neurite branches and E, mean neurite length. Significance levels are indicated as in Figure 1.