Journal: The FASEB Journal

Article Title: ORMDL Proteins Turnover via Proteasome and Autophagy Is Cell‐Type Dependent and Tied to Ceramide Homeostasis

doi: 10.1096/fj.202502924RR

Figure Lengend Snippet: Regulation of ORMDLs stability in human RPE‐1 cells and mouse BMMCs. (A) Levels of ORMDLs, SPTLC1, SPTLC2, and total d18:1 ceramides (measured by LC‐ESI‐MS/MS) were assessed in human RPE‐1 cells and mouse BMMCs after 24‐h treatment with 10 μM myriocin or 10 μM FB 1 , and compared with untreated controls. (B) Quantification of ORMDLs, SPTLC1, SPTLC2, and the LC3‐II/LC3‐I ratio was performed in untreated RPE‐1 cells and BMMCs, and compared with cells treated for 24 h with 50 μM CQ or 10 μM MG132, as determined by immunoblotting. (C) Effects of p97/VCP inhibition by CB‐5083 on ORMDLs, SPTLC1, SPTLC2, the LC3‐II/LC3‐I ratio, ATF4, and p62 levels were assessed in RPE‐1 cells and BMMCs. Protein levels in (A‐C) were normalized to total protein loading using Ponceau S staining. Data are presented as geometric mean ± GSEM ( N = 4). Statistical analysis was performed using one‐way ANOVA with Tukey's post hoc test. Significance levels: p < 0.05 (*), p < 0.01 (**), p < 0.001 (***), p < 0.0001 (****); n.s., not significant.

Article Snippet: Sphingolipid concentrations of S1P, dhC1P, and C1P were determined by single‐point calibration using external standards [ ]: 50 ng S1P d18:1 (Cat# 860492), 50 ng dhC1P d18:0/C16:0 (Cat# 860522), and 50 ng C1P d18:1/C16:0 (Cat# 860533), all from Avanti Polar Lipids.

Techniques: Tandem Mass Spectroscopy, Western Blot, Inhibition, Staining

Journal: Biomolecules

Article Title: Sperm Lipid Composition in Early Diverged Fish Species: Internal vs. External Mode of Fertilization.

doi: 10.3390/biom10020172

Figure Lengend Snippet: Figure 4. High performance thin-layer chromatography (HPTLC)-electrospray ionization ion trap (ESI-IT) mass spectrometry of the unknown spots of the lipid fraction of sterlet sperm. The chromatographic separation of the lipid extract and the corresponding mass spectra of two unknown spots in the positive and in the negative ion mode, respectively, are shown in (a). To further elucidate the structure of m/z 722, this ion was subsequently fragmented by collision-induced decay (b). Due to characteristic fragments with the loss of a hexose moiety (∆= 162 and 180) m/z 722 was assigned to a neutral glycosphingolipid Hex-Cer(d18:1/16:0). Hex: hexose, Cer: ceramide. * not assigned.

Article Snippet: C16 galactosyl(α) ceramide (d18:1/16:0) (Avanti Polar Lipids, Inc., Alabaster, AL, USA) was used as reference.

Techniques: High Performance Thin Layer Chromatography, Mass Spectrometry

Journal: Biomolecules

Article Title: Sperm Lipid Composition in Early Diverged Fish Species: Internal vs. External Mode of Fertilization.

doi: 10.3390/biom10020172

Figure Lengend Snippet: Figure 5. HPTLC-ESI-IT MS of unknown spots of the lipid fraction of stingray sperm. The chromatographic separation of the lipid extract and the corresponding positive and negative ion mass spectra of two unidentified spots are shown in (a). To further elucidate the structure of m/z 824 in the positive and m/z 778 in the negative ion mode, these ions were fragmented by collision-induced decay (b). Due to the detectability in the positive and the negative ion mode and the characteristic fragments with the loss of a sulfate group (∆= 80) and the detection of a sulfated sugar moiety in the negative ion mode (m/z 259 and 241), both signals were assigned to an acidic glycosphingolipid sulpho-Gal-Cer(d18:1/16:0). The signals at m/z 838/792, 840/794, and 854/808 could be putatively assigned to sulpho-Gal-Cer(d18:1/17:0), sulpho-Gal-Cer(d18:0/17:0), and sulpho-Gal-Cer(d18:0/18:0), respectively. Gal—galactose, Cer—ceramide. The assignment of the hexose unit to galactose was made due to the characteristic biosynthesis pathway of sulfo glycolipids [51]. * not assigned.

Article Snippet: C16 galactosyl(α) ceramide (d18:1/16:0) (Avanti Polar Lipids, Inc., Alabaster, AL, USA) was used as reference.

Techniques: High Performance Thin Layer Chromatography