Targeting PLD1, DGKζ, and DGKα in cells

For PLD1 targeting, previously described oligos were used,^{50 (link)} and knockdown efficiency was assessed.^{51 (link)} For DGKζ targeting, we used the previously validated sequences 5′-CUAUGUGACUGAAGAUCGCATT-3′ and 5′-GGUGAAGA GCUGAUUGAGGTT-3′.^{22 (link), 38 (link), 52 (link)} DGKα was silenced with validated sequences;^{32 (link)} either a scrambled (Ambion, Austin, TX, USA) or the equivalent mouse sequence was used as control. For transient targeting, distinct siRNAs were transfected in cells using Oligofectamine (Invitrogen, Carlsbad, CA, USA). For stable targeting, sequences that target murine (control) or human DGKζ were cloned in the pSuperRetro vector (Oligoengine, Seattle, WA, USA), and SW480 cells stably expressing shRNAi were obtained by infection with pSuperRetro-cloned sequence-containing retroviruses using standard protocols.