Journal: PLoS ONE

Article Title: Influence of Tryptophan Contained in 1-Methyl-Tryptophan on Antimicrobial and Immunoregulatory Functions of Indoleamine 2,3-Dioxygenase

doi: 10.1371/journal.pone.0044797

Figure Lengend Snippet: Sources of used substances.

Article Snippet: 1-Methyl-DL-tryptophan , G0411 , 98 , Santa Cruz Biotechnology Inc. (Santa Cruz, USA).

Techniques:

Journal: JCI Insight

Article Title: Interleukin-13 disrupts type 2 pneumocyte stem cell activity

doi: 10.1172/jci.insight.131232

Figure Lengend Snippet: (A) Constitutive overexpression of IL-13 from airway epithelial cells in the Scgb1a1-Il-13–transgenic mouse leads to airspace enlargement and a proportional increase in the number of AEC2s (labeled with DC-LAMP [LAMP3]) compared with AEC1s (marked by podoplanin [PDPN] expression). (B) The proportion of AEC2s to AEC1s was assessed by counting AEC2s (DC-LAMP+ cells) and AEC1s (cells with nuclear expression of homeobox only protein X, HOPX) and expressing the results as a proportion (AEC2/AEC1). There are more AEC2s relative to AEC1s in the IL-13–overexpressing mice. Unpaired t test; error bars indicate mean ± SD. (C) Schematic for lineage-labeling AEC2s in adult mice with (SftpcTmIL13) and without (SftpcTm) constitutive overexpression of IL-13 and subsequent pneumonectomy procedure (PNX). (D) Control lungs 14 days after PNX contain many lineage-labeled AEC1s. (E) IL-13–overexpressing lungs (IL-13 PNX) contain fewer lineage-labeled AEC1s after PNX when compared with controls (control PNX) despite higher levels of post-PNX AEC2 proliferation (data not shown and Supplemental Figure 1). Unpaired t test; error bars indicate mean ± SD. Scale bars: 100 μm (A), 75 μm (D). **P < 0.005.

Article Snippet: Antibodies were as follows: rat, DC-LAMP/CD208 (1:250; DDX0191; Dendritics); rabbit, SFTPC (1:1000; ab3786; Abcam); rabbit, HOPX (1:250; SC-30216; Santa Cruz Biotechnology); rabbit, keratin 5 (1:1000; PRB-160P; Covance); hamster, PDPN (T1-α) (1:1000; clone 8.1.1; DSHB); rabbit, RFP (1:250; 600–401379; Rockland); goat, SCGB1A1 (CC10) (1:200; gift from Barry Stripp, Cedars Sinai, Los Angeles, California, USA); guinea pig, LPCAT (1:2000, gift from John Shannon, Cincinnati Children’s Hospital, Cincinnati, Ohio, USA); rat, KI-67 (1:250; 14-5698-82; Invitrogen, Thermo Fisher Scientific); chicken, GFP (1:500; GFP1020; Aves Lab); rabbit, SOX9 (1:1000, ab53121; Abcam); mouse, p63 (1:200; 58033; One World Lab); rat, RAGE (1:200, MAB1179, R&D Systems); rabbit, AQP5 (1:200, ab78486, Abcam); rabbit, ABCA3 (1:250, WRAB-ABCA3, Seven Hills); mouse, MUC-5ac (1:200, ab77995, Abcam); mouse, α-SMA (1:250, A2547, MilliporeSigma); and human, HTII-280 (1:60, gift from Leland Dobbs, UCSF, San Francisco, California, USA).

Techniques: Over Expression, Transgenic Assay, Labeling, Expressing

Journal: JCI Insight

Article Title: Interleukin-13 disrupts type 2 pneumocyte stem cell activity

doi: 10.1172/jci.insight.131232

Figure Lengend Snippet: (A) Control organoids express both AEC2 (DC-LAMP) and AEC1 (HOPX, PDPN) markers. The morphology of most IL-13–treated spheres is abnormal, with only a single layer of epithelial cells. There are AEC2 and AEC1 markers present, but these occur in an apparently random distribution not seen in control spheres. (B) Compared with control organoids, IL-13–treated organoids contain a higher proportion of cells that lack both AEC2 and AEC1 markers. One-way ANOVA; error bars indicate mean ± SD. **P < 0.005. (C) RNA-Seq of day 16 organoids grown in the presence of IL-13 (compared with control conditions) reveals increased expression of bronchiolar markers (n = 3 biological replicates per condition; asterisk indicates genes significantly differentially expressed). (D) Lineage-labeled (Tomato+) organoids grown in the presence of IL-13 express more SOX9 and display more proliferation (MKI67) than lineage-labeled organoids grown in control conditions. (E) IL-13 induces ectopic expression of KRT5 in AEC2s. An AEC2-derived sphere (arrow) grown in control conditions expresses AEC2 (DC-LAMP) and AEC1 (PDPN) markers but not KRT5. A rare contaminating basal cell–derived organoid (asterisk) serves as a positive control, demonstrating expected KRT5 staining. In contrast, a proportion of IL-13–treated spheres express KRT5. (F) KRT5 expression in IL-13–treated spheres is supported with use of a Krt5GFP reporter mouse line. Scale bars: 50 μm (A); 50 μm, insets 25 μm (D); 75 μm (E); and 50 μm (F).

Article Snippet: Antibodies were as follows: rat, DC-LAMP/CD208 (1:250; DDX0191; Dendritics); rabbit, SFTPC (1:1000; ab3786; Abcam); rabbit, HOPX (1:250; SC-30216; Santa Cruz Biotechnology); rabbit, keratin 5 (1:1000; PRB-160P; Covance); hamster, PDPN (T1-α) (1:1000; clone 8.1.1; DSHB); rabbit, RFP (1:250; 600–401379; Rockland); goat, SCGB1A1 (CC10) (1:200; gift from Barry Stripp, Cedars Sinai, Los Angeles, California, USA); guinea pig, LPCAT (1:2000, gift from John Shannon, Cincinnati Children’s Hospital, Cincinnati, Ohio, USA); rat, KI-67 (1:250; 14-5698-82; Invitrogen, Thermo Fisher Scientific); chicken, GFP (1:500; GFP1020; Aves Lab); rabbit, SOX9 (1:1000, ab53121; Abcam); mouse, p63 (1:200; 58033; One World Lab); rat, RAGE (1:200, MAB1179, R&D Systems); rabbit, AQP5 (1:200, ab78486, Abcam); rabbit, ABCA3 (1:250, WRAB-ABCA3, Seven Hills); mouse, MUC-5ac (1:200, ab77995, Abcam); mouse, α-SMA (1:250, A2547, MilliporeSigma); and human, HTII-280 (1:60, gift from Leland Dobbs, UCSF, San Francisco, California, USA).

Techniques: RNA Sequencing Assay, Expressing, Labeling, Derivative Assay, Positive Control, Staining