Fig. 4. Members of the PLA2G4 and PNLIP family promote BMP synthesis independently of CLN5.

A, B Basal LPG and BMP content in WT and CLN5-KO cells (n = 3 biological replicates). C, D LPG and BMP content of WT and CLN5-KO cells after supplementation with di-oleoyl PG (50 µM) for 6 h (n = 3 biological replicates). E PG-induced increase in BMP levels in CLN5-KO and WT cells (n = 3). F, G BMP content of CLN5-KO cells upon overexpression of indicated PLA2G4 and PNLIP family members under basal conditions and after 8 h PG supplementation (50 µM, n = 3 biological replicates). The WT control was transfected with empty vector. Note that cells used for experiments A–G were pre-cultivated in DMEM containing 10% FBS. For experiments (6-8 h), FBS was replaced by hiFBS to avoid serum-mediated acylation reactions. H, I LPG and BMP content of WT and CLN5-KO cells cultivated for two weeks in DMEM medium containing either 10% FBS or hiFBS (n = 3 biological replicates). J, K LPG and BMP content of WT and CLN5-KO cells cultured in DMEM with 10% hiFBS for one week and subsequently in DMEM with 10% FBS for another week (n = 3 biological replicates). L Secretion of LPG by WT and CLN5-KO HEK293 cells (n = 3 biological replicates). Cells were incubated in DMEM containing 2% BSA for 24 h and lipids were extracted from lyophilized conditioned media. Data are presented as mean ± SD and representative of at least two independent experiments. Statistically significant differences were determined by two-tailed unpaired t-test (B-E, L) without corrections for multiple comparisons, and one-way ANOVA (F, G) followed by corrections for multiple comparisons versus control using Bonferroni post hoc test. Source data are provided as a Source Data file.