Additional file 4: of Network approach identifies Pacer as an autophagy protein involved in ALS pathogenesis

Figure S2. Pacer is expressed in neurons in the spinal cord of wild-type mice a, mRNA levels of Pacer in NSC34 cells depleted of Pacer using shRNA constructs (shRNA A and shRNA B) compared to a scrambled control (shCtrl) were determined by real-time. 18s rRNA levels were used for normalization. Statistical analyses were performed using Student’s t-test. Mean, and SEM with only statistically significant p-values are shown: ***, p ≤ 0.001. b, mRNA levels of Pacer, Rubicon and Beclin1 were determined by quantitative PCR in the spinal cord, cortex, hippocampus, cerebellum, muscle, and liver of wild-type C57BL/6 mice (n=8, 4 females, 4 males). mRNA levels in the liver are used as a reference. c, Confocal microscopy of lumbar spinal cord sections of wild-type mice. Z-stack of confocal images, detection of Pacer, the neuron marker NeuN, or the astrocytic marker GFAP, and DAPI detection by immunofluorescence in C57BL/6 46 mice. Scale bars: 300 μm, and 20 μm. Doted inset indicates where higher magnification images were taken. (PPTX 971 kb