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. Author manuscript; available in PMC: 2021 Sep 17.
Published in final edited form as: Cell Chem Biol. 2020 May 12;27(9):1130–1139.e4. doi: 10.1016/j.chembiol.2020.04.009

Fig. 3. Lipoproteins are PM20D1 co-activators in vitro and in vivo.

Fig. 3.

(a) C20:4-Gly hydrolysis activity of mammalian recombinant, purified mouse PM20D1 (0.1 ug/reaction) alone or following incubation with APOB+ or APOA1+ lipoproteins (LDL and HDL, respectively) isolated from PM20D1-KO mice. N=5/group.

(b-d) Protein concentrations (b), apolipoprotein levels (c), and PM20D1 activity as measured by C20:4-Gly hydrolysis activity (d) of fractionated mouse plasma from either WT (blue trace) or APOE-KO (orange trace) mice.

(e) Total plasma N-acyl amino acid levels from WT (blue) or APOE-KO (orange) mice. N=5/group.

(f,g) Representative LC-MS traces of C18:1-Phe (f) and C18:1-Leu/Ile (g) from WT (blue) or APOE-KO (orange) mice.

For (a), LDL or HDL was isolated from plasma of five pooled PM20D1-KO mice. For (b-d), plasma from five mice per group was pooled together and separated by fast protein liquid chromatography-size exclusion (FPLC-SEC). For (e), N=5/group.

For (a), (d), and (e), Student’s two-tailed t-test was used for the indicated comparison or for WT versus APOE-KO mice. * P < 0.05, ** P < 0.01. Data are shown as means ± SEM. See also Table S3 and S4.