Journal: Cell Host & Microbe

Article Title: Epithelial-Cell-Derived Phospholipase A 2 Group 1B Is an Endogenous Anthelmintic

doi: 10.1016/j.chom.2017.09.006

Figure Lengend Snippet: PLA 2 g1B Has Direct Anthelmintic Properties (A) ATP concentration of H.p. L3 larvae after 24 hr treatment with PLA 2 g1B, n = 3. (B) Number of H.p. larvae imbedded in the small intestinal wall 5 days after infection following 24 hr treatment with PLA 2 g1B, n = 5. (C) Luminal H.p. worms in the small intestine 14 days after 1° infection following 24 hr treatment with PLA 2 g1B, n = 10 (data pooled from two independent experiments). (D) Pla2g1b − /− or WT mice were orally infected with 200 L3 H.p. larvae on day 0 and were drug treated (Rx) on days 14 and 15. Mice were then reinfected with PLA 2 g1B-treated L3 H.p. larvae on day 35 and harvested 14 days after infection (2°). Another cohort of Pla2g1b − /− or WT mice were orally infected with 200 L3 H.p. larvae on day 35 and harvested 14 days after infection (1°). (E) Luminal H.p. worms in the small intestine 14 days after 1° or 2° infection following 24 hr treatment with PLA 2 g1B. (F) Luminal H.p. worms in the small intestine 14 days after 1° infection following 24 hr treatment with PLA 2 g1B. (G) Luminal H.p. worms in the small intestine 14 days after 2° infection following 24 hr treatment with PLA 2 g1B. Data are represented as mean ± SEM, n = 4–5. All data re representative of at least two independent experiments. ∗ = p < 0.05, ∗∗∗ = p < 0.001, ∗∗∗∗ = p < 0.0001 determined using a two-way ANOVA with Sidak’s multiple comparison analysis, one-way ANOVA with Dunnett’s multiple comparison analysis, or an unpaired two-tailed t test. See also Figure S6 .

Article Snippet: 8000 exsheathed L3 H. polygyrus larvae were treated with recombinant mouse PLA 2 g1B (Elabscience), recombinant mouse PLA 2 g1B plus manoalide (200 ng/μL final concentration, Santa Cruz Biotechnology) or heat inactivated recombinant mouse PLA 2 g1B in 1mL EnzChek ® PLA 2 reaction buffer (Invitrogen) at room temperature for 24 hr.

Techniques: Concentration Assay, Infection, Comparison, Two Tailed Test

Journal: Cell Host & Microbe

Article Title: Epithelial-Cell-Derived Phospholipase A 2 Group 1B Is an Endogenous Anthelmintic

doi: 10.1016/j.chom.2017.09.006

Figure Lengend Snippet: Pla2g1b-Treatment Related Changes in Lipid Abundance Relative abundances of phosphatidylethanolamine (PE) and other lipids extracted from PLA 2 g1B-treated (10 ng/μL) and control-treated (0 ng/μL) larvae. (A) Identified PEs. Features were regarded as “identified” by comparison of their precursor ion and MS/MS fragments with the LipidBlast library, as outlined in Figure S1 . The arrangement of fatty acid moieties on the glycerol backbone (i.e., whether in the sn- 1 or sn- 2 position) and the position of double bonds could not be inferred. PE 38:3 and PE 38:4 were detected as a mixture of different fatty acid moieties. (B) Putatively identified (annotated) PEs. The features could be “annotated” as PEs by comparison of peak retention time and inter-cluster mass shifts of 28 Da (CH 2 CH 2 ) and intra-cluster mass shifts of 2 Da (indicative of difference in double bond number [fatty acid saturation]) with other, identified PEs. MS/MS could not be performed due to low abundance. Data are shown as normalized intensities expressed in arbitrary units. Data are represented as mean ± SEM, n = 3. ∗ = p < 0.05. TIC: Total ion current. See also Figure S7 .

Article Snippet: 8000 exsheathed L3 H. polygyrus larvae were treated with recombinant mouse PLA 2 g1B (Elabscience), recombinant mouse PLA 2 g1B plus manoalide (200 ng/μL final concentration, Santa Cruz Biotechnology) or heat inactivated recombinant mouse PLA 2 g1B in 1mL EnzChek ® PLA 2 reaction buffer (Invitrogen) at room temperature for 24 hr.

Techniques: Control, Comparison, Tandem Mass Spectroscopy