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. 2022 Dec 1;64(1):100317. doi: 10.1016/j.jlr.2022.100317

Fig. 4.

Fig. 4

L168F ELOVL4 mutant enhances biosynthesis of VLC-PUFA with carbon chains greater than C34. A: Long-chain PUFA levels in HEK293 cells supplemented with 20:5n3 for 72 h in cells overexpressing L168F ELOVL4 compared with WT ELOVL4 and control (GFP and untransduced [UT]) cells. B: Relative mole % of VLC-PUFA levels normalized to WT ELOVL4 and L168F ELOVL4 protein levels after 20:5n3 supplementation. C: Uptake of 34:5n3 after 72 h in HEK293T cells overexpressing WT, L168F, and control cells (GFP and UT) supplemented with 34:5n3. D: Elongated products of 34:5n3 normalized to WT ELOVL4 and L168F ELOVL4 expression levels. E: WT and L168F ELOVL4 protein expression after 20:5n3 supplementation. Cell lysates were collected after 72 h for Western blot analysis with beta-tubulin as loading control. F: WT and L168F ELOVL4 protein expression after 34:5n3 supplementation. Cell lysates were collected after 72 h for Western blot analysis with beta-tubulin as loading control. Results are the mean ± SD (n = 3). Statistical significance was assessed for A and C by ANOVA with Tukey’s post hoc test and B and D by ANOVA with Šídák’s post hoc test. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001, ns not significant in comparison with WT. ELOVL, elongation of very long-chain fatty acid; UT, untransduced; VLC-PUFA, very long chain-PUFA.