Journal: Cells

Article Title: Palmitic and Stearic Acids Inhibit Chaperone-Mediated Autophagy (CMA) in POMC-like Neurons In Vitro

doi: 10.3390/cells11060920

Figure Lengend Snippet: Analysis of putative CMA protein substrates in lysosomes from POMC-like neurons treated with PA and SA. POMC-like neurons (N43/5) were incubated with heavy DMEM/FBS mixture (SILAC Lys6 Arg6) and then incubated with 100 mM of PA, SA, or vehicle (BSA) for 6 h. Lysosomal fractionations were isolated from each condition as described in and their proteomic composition was analyzed by SILAC-based proteomics. The data were filtered using the following parameters: (i) proteins identified with high false discovery rates (FDRs) and with more than one peptide, (ii) proteins with a p -value of 0.05 (or less), and (iii) proteins with a fold change of 2. ( A ) Pie chart showing the number and percentage of up- and downregulated proteins. ( B ) Number and percentage of proteins from A containing KFERQ-like motifs as analyzed using https://rshine.einsteinmed.org (accessed on 9 February 2022). ( C ) The number and percentage of proteins containing KFERQ-like motifs that are up- and downregulated in PA and SA. ( D ) The number and percentage of proteins containing different KFERQ-like motifs.

Article Snippet: Embryonic Mouse Hypothalamic N43/5 cell lines were grown in DMEM (Hyclone SH30243.02 Cyvita, Marlborough, MA, USA) supplemented with 10% fetal bovine serum (GIBCO, Thermo Fisher Scientific, Waltham, MA, USA) and antibiotic-antimycotic solution (GIBCO 15240-062, Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C in a humidified environment with 5% CO 2 .

Techniques: Incubation, Isolation