Extended Data Fig. 8: Tmed2:SL interaction upon CerS1 deficiency.
From: Sphingolipid subtypes differentially control proinsulin processing and systemic glucose homeostasis
a, Double cut CrispR/Cas9 knockout strategy for CerS1 in Ins1E cells. b, Relative mRNA expression of Sgpl1 and various CerS in wildtype Ins1E cells, Sgpl1ΔIns1E and CerS1:Sgpl1ΔIns1E cells. n = 3 independent experiments. Wildtype Ins1E cDNA was also used for qPCR in Supplementary Fig. 3c. c, Quantification of ceramides of various acyl chain lengths in Sgpl1ΔIns1E and CerS1:Sgpl1ΔIns1E cells by targeted lipidomics (n = 4 independent experiments). Lipid amounts were normalized to cell pellet weights. d, pacSph pull-down of endogenous Tmed2 in Sgpl1ΔIns1E and CerS1:Sgpl1ΔIns1E cells (n = 3 independent experiments); exemplary immunoblot (left) and quantification (right). Eluate intensities were normalized to respective input intensities. e, Experimental setup for knock-down of Tmed1, Tmed2 and both in murine pseudoislets. Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparisons test for each individual mRNA target in (b), multiple two-sided t-tests with Holm-Sidak correction in (c) and repeated measures one-way ANOVA with Tukey’s multiple comparisons test in (d). Data points represent independent experiments. Bar graphs represent means + s.e.m. The Sgpl1ΔIns1E and CerS1:Sgpl1ΔIns1E cells are pools of 3 individual monoclonal cell lines, respectively. Source numerical data and unprocessed blots are available in source data.
