Figure 5. Knockdown of FLOT1 lowers Far1 expression.

(A) Expression levels of Far1 and Flot1 were assessed in HeLa cells transfected with control dsRNA (lane 1) or two different dsRNAs (lanes 3 and 4) against FLOT1. Expression level of Far1 was represented by taking as 100 that in mock-treated cell (lower panel). (B) Plasmalogen synthesis was assessed in mock-treated (−) or FLOT1-knocked down (#68 and #69) HeLa cells by metabolically labelling with ¹⁴C-Etn for 2 h (upper panel). Biosynthesis of plasmalogens was represented by taking as 100 that in mock-treated cells (lower panel). *p < 0.05, **p < 0.01, and ***p < 0.001; one-way ANOVA with Dunnett’s post hoc test as compared with mock-treated cells. (C) Left panel, total cellular level of plasmalogens was represented by taking as 100 that in mock-treated cells. Right panel, expression level of FAR1 relative to ribosomal protein 3 (RPL3)⁶³ was determined by real-time PCR using total RNA, and represented by taking as 100 that in HeLa cells (−). n.s., not significant; one-way ANOVA with Dunnett’s post hoc test as compared with mock-treated cells. Data of unprocessed original blots are available in Supplementary Fig. S2.