CM and SM cooperate to regulate multiple cellular functions. (A) Western blot of UPR markers (left panel) and quantification of protein levels (right panel) in AML12 treated with Ctrl/CM for the indicated time. The diamond indicates the nonspecific band. The ratio of protein band signals in lane 3 (Ctrl, 6 h) was set as 1. (B) Measurement of Ca2+ signals in the cytosol of AML12 by Fura-2 staining. Cells were pretreated with Ctrl or CM for 24 h. (C) Oil Red O staining of AML12 cells after 48-h treatment with Ctrl or CM. Scale bar, 50 μm. (D) Cell proliferation of AML12 treated with Ctrl/CM ± SM (100 μg/ml) for 24 h assessed by crystal violet staining, n = 3. *P < 0.05. (E) Cell death percentage of AML12 treated with Ctrl/CM ± SM (100 μg/ml) for 48 h, n = 3. *P < 0.05. (F) Cell death percentage of AML12 treated with Ctrl/CM ± KIRA8 (0.5 μM) for 48 h, n = 3. *P < 0.05. (G) Schematic illustration of how SM/Cer ratio regulates membrane fluidity and related cell functions. All statistical analyses were calculated by one-way ANOVA. Data were shown as the mean ± SEM. Source data are available for this figure: SourceData F9.