Journal: Molecular cell

Article Title: Methyl-Metabolite Depletion Elicits Adaptive Responses to Support Heterochromatin Stability and Epigenetic Persistence

doi: 10.1016/j.molcel.2020.03.004

Figure Lengend Snippet: Data and Software Availability

Article Snippet: Leupeptin Hemisulfate , Thermo Fisher , Cat#AAJ61188MC.

Techniques: Software, Recombinant, Sequencing, Modification, Extraction, Reverse Transcription, Multiplex sample analysis, Plasmid Preparation, SYBR Green Assay, Staining, cDNA Synthesis, DNA Purification, Purification, Control, Targeted Proteomics

Journal: Molecular cell

Article Title: Methyl-Metabolite Depletion Elicits Adaptive Responses to Support Heterochromatin Stability and Epigenetic Persistence

doi: 10.1016/j.molcel.2020.03.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Leupeptin Hemisulfate , Thermo Fisher , Cat#AAJ61188MC.

Techniques: Recombinant, Sequencing, Modification, Extraction, Reverse Transcription, Multiplex sample analysis, Plasmid Preparation, SYBR Green Assay, Staining, cDNA Synthesis, DNA Purification, Purification, Control, Software, Targeted Proteomics

Journal: The Journal of biological chemistry

Article Title: NPC1 plays a role in the trafficking of specific cargo to melanosomes.

doi: 10.1016/j.jbc.2023.105024

Figure Lengend Snippet: Figure 5. NPC1 impairs TYR processing and degradation. A, cells were untreated or treated with bafilomycin A (100 nM, 5 h) and analyzed by Western blotting for TYR N-glycan processing after Endo H and PNGase F glycosidase treatment of cell lysates. Relative expression levels of PNGase F-sensitive TYR and the ratio of Endo H-resistant to Endo H-sensitive TYR were plotted. Quantitative representation as mean ± SD of relative protein expressions and the ratio of Endo H resistant to Endo H sensitive bands (one-way ANOVA analysis; *p < 0.05, **p < 0.01, n = 3 biological replicates). B, visual comparison of cell pellets of MNT-WT and NPC1-KO cells untreated or treated with bafilomycin A1 (100 nM, 5 h). C, immunoblot analysis for TYR, LAMP-2, and CNX in cell lysates and in the enrichment of secreted extracellular vesicles from culture supernatant. D, effect of MG132 (50 μM, 5 h) or inhibitor mix (INH = 14.5 uM leupeptin and 106 uM pepstatin, 5 h) cell treatment on TYR expression, analyzed by Western blotting and densitometry, represented as fold change relative to cells treated with DMSO. Quantitative representation as mean ± SD of relative protein expression, n = 3 biological replicates (one-way ANOVA analysis; *p < 0.05). E, CHX treated MNT-WT and NPC1-KO cells analyzed by immunoblotting for TYR normalized to tubulin and graphic representation of TYR degradation rate as a percentage of control (CHX, 0 h) (n = 2 biological replicates). CHX, cycloheximide; CNX, calnexin; DMSO, dimethyl sulfoxide; Endo H, endoglycosidase H; LAMP, lysosome-associated membrane protein; NPC1, Niemann–Pick type C1; PNGase F, peptide-N glycosidase F; TYR, tyrosinase.

Article Snippet: MG132 (sc-201260), leupeptin hemisulfate (sc295358) and pepstatin A (sc-45036) were from Santa Cruz Biotechnology.

Techniques: Western Blot, Glycoproteomics, Expressing, Comparison, Control, Membrane