Fig. 1

Validation of Rab10 and pRab10 antibodies in the brain using Rab10ASOs C57BL/6JWT mice were intracerebroventricularly injected with control and Rab10 ASO1 to confirm the specificity of the Rab10 and pRab10 antibodies. a Immunoblot of Rab10 in Rab10 ASO (Rab10 ASO1) and control ASO injected brain samples for cortex, midbrain and striatum. Hsc70 was used as a loading control. b Quantitation of Rab10 protein and normalization with the loading control, Hsc70, showed reduction of Rab10 in brain regions from the Rab10 ASO1 injected mouse (gray bar) compared to control ASO (black bar) in the cortex (p value = 0.002), midbrain (p value = 0.04) and in the striatum (p value = 0.06). Nonparametric Mann–Whitney test was run for statistical analyses (n = 6 biologically independent samples). c Immunoblots showed reduction of pRab10 in Rab10 ASO injected brain samples (Rab10 ASO1) compared to control ASO injected in brain samples in the cortex, midbrain and striatum. Hsc70 was used as a loading control. d Quantitation of pRab10 protein and normalization with the loading control, Hsc70, showed reduction of pRab10 in the Rab10 ASO1 injected mouse samples (gray bar) compared to control ASO injected mouse samples (black bar) in the cortex (p value = 0.004), midbrain (p value = 0.002) and in the striatum (p value = 0.004). Nonparametric Mann–Whitney test was run for statistical analyses (n = 6 biologically independent samples). e Immunofluorescence staining for pRab10 (green) in the striatum showed a reduction of pRab10 in the Rab10 ASO injected mice compared to control ASO injected mice. NeuN (red) immunofluorescence is used to help visualize location on neuronal nuclei and comparable staining in the brain area. Scale bar = 500 µm. All images were captured at the same laser power, gain and offset. f The zoom image shows pRab10 (green) reduction in the Rab10 ASO injected sample compared to control ASO sample. Scale bar = 200 µm. g Normalization of pRab10 integrated density immunofluorescence signal with NeuN signal and quantitation using nested independent t-test showed a significant reduction (p value = 0.0008) in the Rab10 ASO injected samples (gray bar) compared to control samples (black bar) (n = 3 biologically independent samples)