Journal: Cells
Article Title: Obatoclax Rescues FUS-ALS Phenotypes in iPSC-Derived Neurons by Inducing Autophagy.
doi: 10.3390/cells12182247
Figure Lengend Snippet: Figure 7. Proteomic analysis of iPSC-derived neurons. (a) Principle component analysis (PCA) plot of the proteome of the samples analyzed: WT FUS-eGFP treated with DMSO, P525L FUS-eGFP treated with DMSO, and P525L FUS-eGFP treated with obatoclax at 10 nM for 24 h. Three independent experiments. See also Tables S1 and S2. (b) Volcano plot of the comparison between WT FUS-eGFP treated with DMSO and P525L FUS-eGFP treated with DMSO. (c) Volcano plot of the comparison be- tween P525L FUS treated with DMSO and P525L FUS-eGFP treated with obatoclax at 10 nM for 24 h. (d) Differentially expressed proteins related to the proteasome pathway. Proteasome subunit alpha type-1 (PSMA1, p = 0.000103), proteasome activator complex subunit 1 (PSME1, p = 0.000251), protea- some activator complex subunit 2 (PSME2, p = 0.000132), and proteasome activator complex subunit 3 (PSME3, p = 0.000054) are downregulated in P525L FUS-eGFP compared with WT FUS-eGFP. (e) Differentially expressed proteins related to the mucin type O-glycan biosynthesis pathway. Lacto- sylceramide alpha-2,3-sialyltransferase (ST3GAL1, p = 0.005826) and beta-1,4-galactosyltransferase 5 (B4GALT5, p = 0.000386) are upregulated in P525L FUS-eGFP treated with obatoclax compared with P525L FUS-eGFP treated with DMSO. (f) Proteins related to endocytic trafficking. EH domain- containing protein 4 (EHD4, 0.000021) and AP-1 complex subunit sigma-2 (AP1S1, p = 0.004056) are upregulated in P525L FUS-eGFP treated with obatoclax compared with P525L FUS-eGFP treated with DMSO. (g) MRG/MORF4L-binding protein (MRGBP, p = 0.000154) related to DNA double-strand break repair. (h) DnaJ homolog subfamily C member 9 (DNAJC9, p = 0.001442) related to histone chaperone network and heat shock-induced response. Related to Tables S1 and S2.
Article Snippet: Subsequently, the primary antibodies mouse anti-tubulin beta 3 (TUBB3) (1:1000, Cat# BLD-801202, Biolegend, San Diego, CA, USA), rabbit cleaved caspase 3 (CC3) (1:400, Cat# 9661S, Cell Signaling), rabbit LC3 (1:1000, Cat# NB600-1384, Novus Biological), and mouse FUS (1:500, Cat# AMAB90549, Sigma) were applied overnight at 4 ◦C in 0.1% BSA in PBS.
Techniques: Derivative Assay, Comparison, Glycoproteomics, Binding Assay