Figure S4.

Related to Figures 3 and 4

Intracellular levels of (A) Arachidonic acid (AA) and (B) PGE2 levels as measured by LC-MS profiling. (C) AA levels from the conditioned media (CM) of indicated cells before and after lipid depletion (LD). (D) Cell proliferation assays of MCF10A PIK3CA WT cells cultured in CM derived from WT or E545K MUT cells before or after LD, with or without the supplementation of 25 μM AA, palmitate or palmitoleate. (E) Cell proliferation assays of MCF10A PIK3CA E545K MUT cells before or after LD, with or without the supplementation of 25 μM AA, palmitate or palmitoleate. (F) Diacylglycerol (DAG) levels in MCF10A PIK3CA WT and MUT cells. (G) Diagram summarizing DAG contribution to AA production. Sulforhodamine B (SRB) protein staining was used in (D) and (E) to measure cell proliferation over 5 days. Data are presented as the mean ± SEM of n = 3-4 biological replicates and are representative of at least two independent experiments. ^∗p ≤ 0.05; ^∗∗p ≤ 0.01; ^∗∗∗p ≤ 0.001. P values in (A), (B), (C) and (F) were calculated using one-way ANOVA followed by unpaired, two-tailed Student’s t test with Bonferroni correction, and in (D) and (E) with two-way ANOVA.