Journal: Leukemia

Article Title: PARP14 is a novel target in STAT6 mutant follicular lymphoma.

doi: 10.1038/s41375-022-01641-x

Figure Lengend Snippet: Fig. 1 STAT6 mutations are associated with activated IL-4 signaling in human FL. A Lolliplot of STAT6 mutations in 258 human FL biopsies. B Unsupervised hierarchical clustering of gene expression data from 106 human FL, annotated for STAT6 genotype. C Same data used for gene set enrichment analysis (GSEA) for established IL-4 signatures comparing STAT6WT and STAT6MUT. D Volcano plot of differentially expressed genes between STAT6WT and STAT6MUT FL (N = 106, log2FC = ±0.65, p value = 0.05). E FCER2 (CD23) gene expression in human FL with wild type STAT6 (STAT6WT) vs any STAT6 mutation (STAT6MUT) vs STAT6 mutations at position D419 (STAT6D419). F Representative immunohistochemistry (IHC) stains for CD23 and pSTAT6 in human FL with STAT6WT (left) and STAT6MUT (STAT6D419G, right).

Article Snippet: OCI-Ly1 OCI-Ly8 EV WT D419G +- +- +-+- R el at iv e Ex pr es si on p≤0.0001 p≤0.0001 150μm 150μm 150μm 150μm - + - +-+ -+ - + - EV WT D419G OCI-Ly1 OCI-Ly8 EV WT D419G + 0 2 4 6 8 IL-4 STAT6 sCD23 (ELISA) 1 2 3 4 0 200 400 600 800 Passage N um be r o f C ol on ie s 0 1000 2000 3000 4000 G eo m et ric M ea n (C D 23 ) 0.0 0.2 0.4 0.6 0.8 WT D419G WT D419G pSTAT6 total STAT6 3xFlag STAT6 Lamin B1 α-Tubulin - + P Cytoplasma Nucleus - + P - + P - + PIL-4 F 97 kDa 97 kDa 97 kDa 68 kDa 50 kDa pSTAT6 total STAT6 3xFlag STAT6 Lamin B1 α-Tubulin IL-4 97 kDa 97 kDa 97 kDa 68 kDa 50 kDa Cytoplasma Nucleus STAT6 STAT6 Leukemia (2022) 36:2281 – 2292 data (nCounter, NanoString) (Fig. 1E).

Techniques: Gene Expression, Mutagenesis, Immunohistochemistry

Journal: Leukemia

Article Title: PARP14 is a novel target in STAT6 mutant follicular lymphoma.

doi: 10.1038/s41375-022-01641-x

Figure Lengend Snippet: Fig. 6 Model of a PARP14-mediated self-reinforcing regulatory circuit that amplifies IL-4 induced transcriptional activity in STAT6MUT FL. STAT6 mutations amplify IL-4 induced STAT6-dependent gene activation via an intracellular self-reinforcing regulatory microcircuit that involves aberrantly increased PARP14 levels in IL-4 stimulated STAT6MUT FL cells. Increased STAT6-dependent gene expression involves cytokines (e.g., CCL17 and CCL22) which contribute to the re-education of the tumor microenvironment, including increased recruitment of IL- 4 producing T follicular helper (TFH) cells. Details see text. Yellow star indicates STAT6 mutation. Figure created with BioRender.com.

Article Snippet: OCI-Ly1 OCI-Ly8 EV WT D419G +- +- +-+- R el at iv e Ex pr es si on p≤0.0001 p≤0.0001 150μm 150μm 150μm 150μm - + - +-+ -+ - + - EV WT D419G OCI-Ly1 OCI-Ly8 EV WT D419G + 0 2 4 6 8 IL-4 STAT6 sCD23 (ELISA) 1 2 3 4 0 200 400 600 800 Passage N um be r o f C ol on ie s 0 1000 2000 3000 4000 G eo m et ric M ea n (C D 23 ) 0.0 0.2 0.4 0.6 0.8 WT D419G WT D419G pSTAT6 total STAT6 3xFlag STAT6 Lamin B1 α-Tubulin - + P Cytoplasma Nucleus - + P - + P - + PIL-4 F 97 kDa 97 kDa 97 kDa 68 kDa 50 kDa pSTAT6 total STAT6 3xFlag STAT6 Lamin B1 α-Tubulin IL-4 97 kDa 97 kDa 97 kDa 68 kDa 50 kDa Cytoplasma Nucleus STAT6 STAT6 Leukemia (2022) 36:2281 – 2292 data (nCounter, NanoString) (Fig. 1E).

Techniques: Activity Assay, Activation Assay, Gene Expression, Mutagenesis

Journal: Scientific Reports

Article Title: A novel peptide ADAM8 inhibitor attenuates bronchial hyperresponsiveness and Th2 cytokine mediated inflammation of murine asthmatic models

doi: 10.1038/srep30451

Figure Lengend Snippet: Lung tissue sections were stained by hematoxylin & eosin (H&E) shown in panel a–c, and immunohistochemistry (IHC) shown in panel d–f to visualize bronchiolar structure and ADAM8 expression, respectively. Panel g-i display zoom-in images corresponding to panel d–f, with arrows pointing to ADAM8-positive eosinophils. The bar in panel a-f equals 25 μm, whereas the bar in panel g-i equals 12.5 μm. The left, middle, and right column of images represents lung tissue sections from control group (panel a, d, g), OVA group (panel b, e, h), and OVA/BK-1361 group (panel c, f, i), respectively. Note the OVA-induced thickening of bronchiolar epithelium compared to control (panel b vs. a), which was reduced by BK-1361 treatment (panel c vs. b), and the OVA-induced fibrotic changes (panel h vs. g), which was also reduced by BK-1361 treatment (panel i vs. h). ADAM8-positive eosinophils (ADAM8 + cells) appeared mainly in OVA group (panel h vs. g), and became much less abundant after BK-1361 treatment (panel i vs. h). Panel j, k displays quantified bronchial wall thickness, and number of ADAM8 + cells, respectively. Data are representative of two independent experiments with n = 30–40 counts from 3–4 age-matched female mice per group. Result are expressed as mean ± SD. **P < 0.005 versus control. ## P < 0.005 versus OVA.

Article Snippet: Co. (Nanjing, China); TRIzol solution was purchased from Bioteke Co. (Beijing, China); RevertAid First Strand cDNA Synthesis Kit was purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA); Oligonucleotide primers were produced by Invitrogen Life Technologies Co. (Shanghai, China); SsoFast EvaGreen Supermix was purchased from Bio-Rad Laboratories (Hercules, CA, USA); ELISA kits for soluble CD23 (sCD23) and soluble ADAM8 (sADAM8) was purchased from either Life Science Inc. (Wuhan, China)or R&D Systems (Wiesbaden, Germany); NanoDrop was purchased from PeqLab (Erlangen, Germany); the ADAM8 antibody (Cat#: orb4376) was purchased from Biorbyt (Cambridge, U.K.), CD23 antibody (abb185807) from Abcam (Cambridge, U.K.), goat anti-mouse HRP and goat anti-rabbit HRP from Jackson ImmunoResearch (West Grove, PA, USA), mouse anti-GAPDH from Thermo Fisher Scientific (Waltham, MA, USA); VECTASTAIN Elite ABC Kit was purchased from Vector Laboratories (Peterborough, U.K.).

Techniques: Staining, Immunohistochemistry, Expressing, Control

Journal: Scientific Reports

Article Title: A novel peptide ADAM8 inhibitor attenuates bronchial hyperresponsiveness and Th2 cytokine mediated inflammation of murine asthmatic models

doi: 10.1038/srep30451

Figure Lengend Snippet: Total CD23 in lung tissue lysates and soluble CD23 (sCD23) in soluble protein fraction detected via ELISA are presented in panel a, and b respectively. The WB analysis results of sCD23 in lung tissue lysates and the representative image ofWB bands of CD23 are displayed in panel c, and d, respectively. Note the band for sCD23 around 38 kDa. For the quantification, values are derived from n = 3 (wild-type) and n = 4 (ADAM8-deficient) and are given as mean ± SEM. *p < 0.05 versus litter treated with saline.

Article Snippet: Co. (Nanjing, China); TRIzol solution was purchased from Bioteke Co. (Beijing, China); RevertAid First Strand cDNA Synthesis Kit was purchased from Thermo Fisher Scientific Inc. (Waltham, MA, USA); Oligonucleotide primers were produced by Invitrogen Life Technologies Co. (Shanghai, China); SsoFast EvaGreen Supermix was purchased from Bio-Rad Laboratories (Hercules, CA, USA); ELISA kits for soluble CD23 (sCD23) and soluble ADAM8 (sADAM8) was purchased from either Life Science Inc. (Wuhan, China)or R&D Systems (Wiesbaden, Germany); NanoDrop was purchased from PeqLab (Erlangen, Germany); the ADAM8 antibody (Cat#: orb4376) was purchased from Biorbyt (Cambridge, U.K.), CD23 antibody (abb185807) from Abcam (Cambridge, U.K.), goat anti-mouse HRP and goat anti-rabbit HRP from Jackson ImmunoResearch (West Grove, PA, USA), mouse anti-GAPDH from Thermo Fisher Scientific (Waltham, MA, USA); VECTASTAIN Elite ABC Kit was purchased from Vector Laboratories (Peterborough, U.K.).

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Saline