c17 glucosyl(ß) ceramide  (Avanti Polar)

Journal: bioRxiv

Article Title: Inhibition of Acid Sphingomyelinase Links Sphingolipid Remodeling to Necroptotic Cell Death

doi: 10.64898/2026.03.10.710852

Figure Lengend Snippet: (A) Heatmap displaying the relative total lipid changes under control, 4 μM ARC39 treatment, necroptosis, and necroptosis with 4 μM ARC39 treatment. The relative abundance is presented as Log 2 (relative abundance). The relative abundance was calculated by normalizing each lipid species to the raw abundance of the internal standard. The internal standard corrected abundances were then normalized to the internal standard-corrected abundance of the respective control. Four biological replicates were analyzed per condition (n = 4). Red indicates accumulation, black indicates baseline levels, and green indicates depletion. Cer, Ceramide; DiHCer, Dihydroceramide; HexCer, Hexosylceramide; SM, Sphingomyelin; DiHSM, Dihydrosphingomyelin; TAG, Triacylglycerol; PC, Phosphatidylcholine. Internal standards used for correction: C17:0 Ceramide for Cer, DiH Cer and Hex Cer; C17:0 Sphingomyelin for SM and DiH SM; C39:0 Triacylglycerol for TAGs. (B) Comparison of the relative total lipids in HT-29 cells across control, 4 μM ARC39, necroptosis, and necroptosis + ARC39 conditions. The relative total lipids were quantified by adding the internal standard-corrected abundance for each lipid species per condition which were then normalized to the total lipids of the control. Data represents the mean ± standard deviation; n = 4, ns represents p > 0.05, * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001.

Article Snippet: C17:0 Ceramide (Cat # 860517P), d Oleic Acid (Cat # 861809), C17:0 Glucosylceramide (Cat # 860569), C17 Sphingomyelin (Cat # 860656P), and C39 TAGs lipid standards were obtained from Avanti Polar Lipids.

Techniques: Control, Comparison, Standard Deviation

Journal: bioRxiv

Article Title: Inhibition of Acid Sphingomyelinase Links Sphingolipid Remodeling to Necroptotic Cell Death

doi: 10.64898/2026.03.10.710852

Figure Lengend Snippet: (A) shSMPD1 knockdown efficiency in HT-29 cells. Short hairpin Red Fluorescent Protein (shRFP) was utilized as a negative control. Relative gene expression was calculated by normalizing SMPD1 expression to the housekeeping gene HPRT1. Data represents the mean ± standard deviation; n = 3, ** represents p < 0.01. (B) Genetic knockdown of SMPD1 increased cell viability compared to the shRFP control under necroptotic conditions in the MTT Assay. Data represents the mean ± standard deviation; n = 5, *** represents p < 0.001. (C) PI uptake decreased following SMPD1 knockdown, suggesting reduced plasma membrane rupture. Data represents the mean ± standard deviation; n = 5, *** represents p < 0.001. (D) Heatmap showing the relative abundance of ceramides in control vs. necroptosis conditions for shRFP and shSMPD1. Relative abundance was calculated by normalizing each ceramide species to the C17 Cer internal standard. and then to its respective control. Data is shown as Log 2 (relative abundance). Internal standard used for correction: C17:0 Ceramide for Cer (E) . Total ceramide levels are reduced upon SMPD1 knockdown. Data represents the mean ± standard deviation; n = 3, ** represents p < 0.01

Article Snippet: C17:0 Ceramide (Cat # 860517P), d Oleic Acid (Cat # 861809), C17:0 Glucosylceramide (Cat # 860569), C17 Sphingomyelin (Cat # 860656P), and C39 TAGs lipid standards were obtained from Avanti Polar Lipids.

Techniques: Knockdown, Negative Control, Gene Expression, Expressing, Standard Deviation, Control, MTT Assay, Clinical Proteomics, Membrane

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: For HEK293 cells, 100 μg of protein was transferred into a 5 mL glass vial preloaded with internal standards: 12.5 ng Sph d17:1 (Cat# 860640), 50 ng dhCer d18:0/C14:0 (Cat# 860632), and 12.5 ng Cer d18:1/C17:0 (Cat# 860517) from Avanti Polar Lipids, Alabaster, AL, USA.

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