Journal: Glia

Article Title: A Human Model of Oligodendrocyte Development Shows MCL ‐1 Influences Oligodendrocyte Morphogenesis

doi: 10.1002/glia.70128

Figure Lengend Snippet: Targeting enzymes of the mitochondrial fatty acid oxidation pathway in a glucose‐depleted state. (A) Representative spinning disk confocal maximum intensity projections of immunofluorescent staining for nuclei (blue), oligodendrocyte transcription factor 2 (OLIG2) (red), platelet‐derived growth factor receptor alpha (PDGFRα) (magenta), and BODIPY (green) in DMSO (vehicle) and etomoxir treated OPCs that were supplemented with BSA‐palmitate saturated fatty acid complex or BSA (vehicle) (scale bar = 25 μm). Zoom‐in of lipid droplets (scale bar = 10 μm). (B) Quantification of lipid droplets per cell in DMSO and etomoxir treated groups supplemented with BSA or BSA‐palmitate. Each color represents a biological replicate ( n = 3), each dot represents a cell (8–12 per n ), each square or triangle represents the mean of the biological replicate, analyzed using an ordinary two‐way ANOVA, error bars represent mean ± SEM. (C) Quantification of lipid droplet volume in DMSO and etomoxir treated groups supplemented with BSA or BSA‐palmitate. Each color represents a biological replicate ( n = 3), each triangle represents the mean of the biological replicate, analyzed by student's t ‐test; error bars represent mean ± SEM. (D) Representative spinning disk confocal maximum intensity projections of immunofluorescent staining for nuclei (blue), oligodendrocyte transcription factor 2 (OLIG2) (red), platelet‐derived growth factor receptor alpha (PDGFRα) (magenta), and BODIPY (green) in DMSO (vehicle) and S63845 treated OPCs that were supplemented with BSA‐palmitate saturated fatty acid complex or BSA (vehicle) (scale bar = 25 μm). Zoom‐in of lipid droplets (scale bar = 10 μm). (E) Quantification of lipid droplets per cell in DMSO and S63845 treated groups supplemented with BSA or BSA‐palmitate. Each color represents a biological replicate ( n = 3), each dot represents a cell (8–12 per n ), each square or triangle represents the mean of the biological replicate, analyzed using an ordinary two‐way ANOVA, error bars represent mean ± SEM. (F) Quantification of lipid droplet volume in DMSO and S63845 treated groups supplemented with BSA or BSA‐palmitate. Each color represents a biological replicate ( n = 3), each triangle represents the mean of the biological replicate, analyzed by student's t ‐test; error bars represent mean ± SEM. (G) Representative spinning disk confocal maximum intensity projections of immunofluorescent staining for nuclei (blue), oligodendrocyte transcription factor 2 (OLIG2) (red), platelet‐derived growth factor receptor alpha (PDGFRα) (magenta), and BODIPY (green) in DMSO (vehicle) and Triacsin C treated OPCs that were supplemented with BSA‐palmitate saturated fatty acid complex or BSA (vehicle) (scale bar = 25 μm). Zoom‐in of lipid droplets (scale bar = 10 μm). (H) Quantification of lipid droplets per cell in DMSO and Triacsin C treated groups supplemented with BSA or BSA‐palmitate. Each color represents a biological replicate ( n = 3), each dot represents a cell (8–12 per n ), each square or triangle represents the mean of the biological replicate, analyzed using an ordinary two‐way ANOVA, error bars represent mean ± SEM. (I) Quantification of lipid droplet volume in DMSO and Triacsin C treated groups supplemented with BSA or BSA‐palmitate. Each color represents a biological replicate ( n = 3), each triangle represents the mean of the biological replicate, analyzed by student's t ‐test; error bars represent mean ± SEM.

Article Snippet: Following sorting, cells were cultured in HBSS (Thermo Fisher Scientific, 14025092) supplemented with 100 μM BSA‐Palmitate Saturated Fatty Acid (Cayman Chemical, 29558) or BSA as the vehicle control (Cayman Chemical, 29556) for 24 h. During the 24‐h period, cells were treated with 50 μM etomoxir (Tocris, 4539), 2 μM S63845 , or 2 μM Triacsin C (Tocris, 2472).

Techniques: Staining, Derivative Assay

Journal: bioRxiv

Article Title: Metabolic imprinting drives epithelial memory during mucosal fungal infection

doi: 10.1101/2025.07.11.664387

Figure Lengend Snippet: A. Intracellular fatty acid concentrations in epithelial cells stimulated without or with β-glucan for 6 and 24h. N=5 ; One-way ANOVA with Dunnett’s multiple comparisons test. B. Intracellular fatty acid concentrations in BG-primed epithelial cells after 48h of rest. N=5 ; Two-tailed Mann– Whitney Test. C. Fatty acids concentrations in epithelial-enriched tissue from immunocompetent wild-type mice. N=6. Two-tailed Mann–Whitney Test. Abbreviations: PA:C16:0 – palmitic acid, POA:C16:1n7 – Palmitoleic acid, SA:C18:0 – Stearic acid, OA: C18:1n9 – oleic acid, α-LA:C18:3n3 – α-linoleic acid and LA:C18:2n6 – linoleic acid. D. Schematic of fatty acid oxidation and TCA cycle. E-G. Representative immunoblot analysis of carnitine palmitoyltransferase (CPT1/2) expression in epithelial cells stimulated without or with β-glucan for the indicated times ( E ), in β-glucan primed epithelial cells after 48h ( F ) and in epithelial-enriched tissue from immunocompetent wild-type mice after fungal clearance with primary infection( G ).

Article Snippet: To identify the metabolic pathways responsible for enhanced cytokines productions, cells were treated with various inhibitors: PRODH inhibitor (10mM, Cat No.185396, sigma), glycolysis inhibitor (10mM, Cat No.S4701, SelleckChem), CPT1 inhibitor (100μm; Cat No. S8244; SelleckChem), histone methylation inhibitor (100μM; Cat No. E1024; SelleckChem), and HIF-1α inhibitor (10μM; IDF-11774; SelleckChem).

Techniques: Two Tailed Test, MANN-WHITNEY, Western Blot, Expressing, Infection