TNF treatment induces accumulation of ceramide and several atypical sphingoid bases (DBS) in MN9D dopaminergic (DA) neuroblastoma cells derived from mouse ventral mesencephalon. MN9D cells were treated with PBS or 5 ng/mL TNF for 24 or 48 hours as indicated prior to cell harvest for lipid extraction as described in Methods. A, Lipidomic analyses indicate time-dependent accumulation of ceramide (Cer), sphingomyelin (SM), and hexosylceramide (HexCer) in TNF-treated cells relative to PBS-treated cells. B, Lipidomic analyses also revealed time-dependent accumulation of sphingosine (So), sphinganine (Sa), sphingosine-phosphate (SoP), sphinganine-phosphate (SaP), 1-deoxysphinganine (1-deoxySa), and 1-desoxymethylsphinganine (1-desoxyMeSa) after treatment with TNF. All values represent group means +/− SEM, n = 3. One-way ANOVA with Tukey’s post-hoc test; *** denotes p < 0.001 and * denotes p < 0.05 compared to PBS at 48 hrs. N.S. denotes not significant.