Fig. 8.

CMIP induces alterations in the glycosphingolipid biosynthetic pathway and ganglioside species. A mouse podocyte cell line was stably transfected with a CMIP expression vector or an empty vector (EV). Lipids were extracted and subjected to LC-MS analysis. a Principal component analysis of ion abundance in the positive and negative modes discriminated between CMIP-transfected and EV-transfected cells. PC1 accounted for 74.6% of the total variance. b Metabolome view after MetPA shows sphingolipid metabolism to be the pathway most significantly altered by CMIP expression. Total: the total number of compounds in the pathway; Hits: the actual matched number from the uploaded data; Raw p: original p value calculated from enrichment analysis; Holm p: p value adjusted by the Holm-Bonferroni method; FDR p: p value adjusted using the false discovery rate; Impact: pathway impact value calculated from pathway topology analysis. c The relative abundances of several ganglioside species are very significantly different between empty vector-transfected and CMIP-transfected cells. Data are expressed as the means ± SDs of normalized arbitrary units. ***p < 0.001. EV: n = 3. CMIP: n = 6. d Representative Western blot showing GM3 synthase (St3Gal5 gene) in protein lysates at rest (0 min) or after 30 and 60 min of the activation of transgenic and WT T cells with anti-CD3/CD28 antibodies; blots were stripped and reprobed with anti-GAPDH antibody. Statistical analyses of three independent experiments were performed [Tg vs. WT (30 min), ***p = 0.0005 and Tg vs. WT (60 min), *p = 0.0254)]