Journal: PLOS One

Article Title: TLR9/MyD88/NF-κB signaling mediates mental stress-induced exacerbation of psoriasis through immune dysregulation in a mouse model

doi: 10.1371/journal.pone.0344474

Figure Lengend Snippet: (A-F): qRT-PCR was used to detect the expression level of TLR9, MyD88, NF-κB p65, TGF-β, IL-10 and IL-17 mRNA; (G-I): The concentrations of TGF-β, IL-10 and IL-17 were detected by ELISA. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The samples were then incubated with a primary antibody targeting p-NF-κB p65 (Proteintech), IL-1β (Proteintech), IL-6 (Abcam), IL-17 (Proteintech) and IL-23 (Abcam) (all at a dilution of 1:50).

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: PLOS One

Article Title: TLR9/MyD88/NF-κB signaling mediates mental stress-induced exacerbation of psoriasis through immune dysregulation in a mouse model

doi: 10.1371/journal.pone.0344474

Figure Lengend Snippet: (A) : The images of the affected skin in the different groups were photographed. (B) : Sucrose preference rate and stationary time were assessed by sucrose preference test and tail suspension test respectively. (C) : HE analysis of skin tissue in each group, scale bar = 50 μm; (D-F) : The expression levels of TLR9, MyD88, and NF-κB p65 in PBMCs of mice were detected by qRT-PCR; (G) The fluorescence intensity of p-NF-κB p65 was assessed by immunofluorescence test, scale bar = 100 μm; (H) The expression levels of TGF-β, IFN-γ, IL-1β, IL-6, IL-17 and IL-23 in PBMCs of mice were detected by qRT-PCR; (I) The fluorescence intensities of IL-1β, IL-6, IL-17 and IL-23 were assessed by immunofluorescence test, scale bar = 100 μm. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The samples were then incubated with a primary antibody targeting p-NF-κB p65 (Proteintech), IL-1β (Proteintech), IL-6 (Abcam), IL-17 (Proteintech) and IL-23 (Abcam) (all at a dilution of 1:50).

Techniques: Suspension, Expressing, Quantitative RT-PCR, Fluorescence, Immunofluorescence

Journal: PLOS One

Article Title: TLR9/MyD88/NF-κB signaling mediates mental stress-induced exacerbation of psoriasis through immune dysregulation in a mouse model

doi: 10.1371/journal.pone.0344474

Figure Lengend Snippet: Following treatment with the specific inhibitors of the TLR9/MyD88/NF-κB pathway, (A): The protein levels of TLR9, MyD88, p-NF-κB p65 and NF-κB p65 in PBMCs of mice were measured by western blotting; (B) The fluorescence intensity of p-NF-κB p65 was assessed by immunofluorescence test, scale bar = 100 μm; (C): The serum concentrations of TGF-β, IFN-γ, IL-1β, IL-6, IL-17 and IL-23 were detected by ELISA; (D) The fluorescence intensities of IL-1β, IL-6, IL-17 and IL-23 were assessed by immunofluorescence test, scale bar = 100 μm. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The samples were then incubated with a primary antibody targeting p-NF-κB p65 (Proteintech), IL-1β (Proteintech), IL-6 (Abcam), IL-17 (Proteintech) and IL-23 (Abcam) (all at a dilution of 1:50).

Techniques: Western Blot, Fluorescence, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: PLOS One

Article Title: TLR9/MyD88/NF-κB signaling mediates mental stress-induced exacerbation of psoriasis through immune dysregulation in a mouse model

doi: 10.1371/journal.pone.0344474

Figure Lengend Snippet: Following injection of LV-NC, LV-shTLR9, or LV-MyD88 in mice, (A) : Daily scores based on PASI. (B) : HE analysis of skin tissue in each group, scale bar = 50 μm; (C-E) : qRT-PCR was used to detect the expression level of TLR9, MyD88, and NF-κB p65 mRNA in PBMCs of mice; (F) The fluorescence intensity of p-NF-κB p65 was assessed by immunofluorescence test, scale bar = 100 μm; (G) : The concentrations of TGF-β, IFN-γ, IL-1β, IL-6, IL-17 and IL-23 were detected by ELISA; (H) The fluorescence intensities of IL-1β, IL-6, IL-17 and IL-23 were assessed by immunofluorescence test, scale bar = 100 μm. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The samples were then incubated with a primary antibody targeting p-NF-κB p65 (Proteintech), IL-1β (Proteintech), IL-6 (Abcam), IL-17 (Proteintech) and IL-23 (Abcam) (all at a dilution of 1:50).

Techniques: Injection, Quantitative RT-PCR, Expressing, Fluorescence, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Obstructive Sleep Apnea–Induced Hypertension Is Associated With Increased Gut and Neuroinflammation

doi: 10.1161/JAHA.122.029218

Figure Lengend Snippet: Plasma levels of interleukin‐17a ( A ) and interleukin‐10 ( B ). Systolic blood pressure ( C ) in sham and OSA rats treated with a nonimmune IgG or interleukin‐17a neutralizing antibody. n=5 in ( A and B ), n=5–6 in ( C ); * P <0.05 for Sham+IgG vs OSA+IgG, # P <0.05 of OSA+nIL‐17a vs OSA+IgG, *** P <0.001 using two‐way ANOVA ( A , B ) or 2‐way repeated measures ANOVA and multiple comparisons with Tukey's multiple comparisons test ( C ). Error bars represent ±SEM. BP indicates blood pressure; IgG, immunoglobulin G; and OSA, obstructive sleep apnea.

Article Snippet: Biotinylated anti‐rat interleukin‐17a (Rockland, No. 212‐406‐b32) was added and incubated for 2 hours at room temperature.

Techniques: Clinical Proteomics

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Obstructive Sleep Apnea–Induced Hypertension Is Associated With Increased Gut and Neuroinflammation

doi: 10.1161/JAHA.122.029218

Figure Lengend Snippet: Effects of interleukin‐17a neutralization on Tregs, T H 1, T H 17, TNFα+, and macrophages in the ileum ( A ), cecum ( B ), and brain ( C ) of sham and OSA rats. n=4 in ileum, n=6 in cecum and brain; * P <0.05, ** P <0.005, *** P <0.0005, **** P <0.0001 using 2‐way ANOVA with Tukey's multiple comparisons test. Error bars represent ±SEM. OSA indicates obstructive sleep apnea; T H , T helper cell; and Tregs, T regulatory cells.

Article Snippet: Biotinylated anti‐rat interleukin‐17a (Rockland, No. 212‐406‐b32) was added and incubated for 2 hours at room temperature.

Techniques: Neutralization

Journal: Investigative Ophthalmology & Visual Science

Article Title: METTL3 Potentiates Corneal Allograft Rejection via Increased Pro-Inflammatory Cytokine Production and Aerobic Glycolysis

doi: 10.1167/iovs.67.2.48

Figure Lengend Snippet: Inhibition of METTL3 activity significantly attenuates the immune rejection response in corneal transplantation. ( A ) Experimental protocol for subconjunctival injection of STM2457/Vehicle in the treatment of allogeneic corneal transplantation rejection. ( B ) Representative images of different treatment groups following allogeneic corneal transplantation on postoperative day 20. ( C ) The survival curves of allogeneic corneal transplants in each group were analyzed using the Kaplan-Meier method. ( D ) Protein levels of IL-17A and TNF-α in corneal grafts subjected to different treatments were determined by ELISA. ( E ) RT-qPCR was used to assess the transcriptional levels of Il1b , Il6 , Il12a , and Il17a in corneal allografts across different groups. ( F ) Lactate levels in corneal allografts from different groups were measured using a Lactate Kit. ( G ) The ECAR measurements were obtained following the sequential addition of glucose (10 mM), oligomycin (1.5 µM), and 2-DG (50 mM) at the indicated time points. ( H, I ) Representative Western blot H and quantitative analysis I of glycolysis-related proteins (HIF-1α, MCT-4, and LDHA) in corneal grafts from mice subjected to different treatments. Uncropped immunoblots are shown in A. ( J, K ) Phosphorylation levels of S6K, AKT, and 4E-BP1 in corneal grafts from different groups were detected by Western blot J and quantitatively analyzed K . Uncropped immunoblots are shown in B. Nor, normal group; Vehicle, the allogeneic group topically treated with vehicle (excipient solution); STM2457, the allogeneic group topically treated with STM2457. * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant.

Article Snippet: The concentrations of IL-6, IL-17A, and tumor necrosis factor-α (TNF-α) in the supernatant of stimulated BMDCs and corneal samples were quantified using commercial mouse enzyme-linked immunosorbent assay (ELISA) kits (KE10091, KE10020, and KE10002; Proteintech, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Inhibition, Activity Assay, Transplantation Assay, Injection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Western Blot, Phospho-proteomics

Journal: Investigative Ophthalmology & Visual Science

Article Title: METTL3 Potentiates Corneal Allograft Rejection via Increased Pro-Inflammatory Cytokine Production and Aerobic Glycolysis

doi: 10.1167/iovs.67.2.48

Figure Lengend Snippet: Inhibition of METTL3 activity significantly reduces the m 6 A levels and inflammatory responses in BMDCs. ( A ) The m 6 A levels in BMDCs of different groups. ( B ) The protein levels of IL-6 and TNF-α in BMDCs from different groups were determined by ELISA. ( C ) The transcriptional levels of Il1b , Il6 , Il12a , and Il17a in BMDCs from different groups were assessed by RT-qPCR. CON, untreated group; LPS, cells were treated with LPS (200 ng/mL) group; LPS + STM2457, cells were treated with LPS (200 ng/mL) and STM2457 (100 nM) group. ** P < 0.01, *** P < 0.001; ns, not significant.

Article Snippet: The concentrations of IL-6, IL-17A, and tumor necrosis factor-α (TNF-α) in the supernatant of stimulated BMDCs and corneal samples were quantified using commercial mouse enzyme-linked immunosorbent assay (ELISA) kits (KE10091, KE10020, and KE10002; Proteintech, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Inhibition, Activity Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Investigative Ophthalmology & Visual Science

Article Title: METTL3 Potentiates Corneal Allograft Rejection via Increased Pro-Inflammatory Cytokine Production and Aerobic Glycolysis

doi: 10.1167/iovs.67.2.48

Figure Lengend Snippet: Genetic knockdown of METTL3 significantly inhibits the inflammatory response and glycolytic activity of BMDCs. ( A ) The m 6 A content in BMDCs from different groups was quantified using a colorimetric m 6 A assay. ( B ) The protein levels of IL-6 and TNF-α in BMDCs from different groups were determined by ELISA. ( C, D ) Western blotting was used to detect the expression levels of glycolysis-related proteins (HIF-1α, MCT-4, and LDHA) in BMDCs from different groups C , and the results were quantified using Image J software D . Uncropped immunoblots are shown in A. ( E, F ) Western blotting was performed to assess the protein levels of METTL3 and the phosphorylation levels of S6K, AKT, and 4E-BP1 in BMDCs subjected to different treatments E , with subsequent quantification analysis conducted using Image J software F . Uncropped immunoblots are shown in B. CON, untreated group; LPS, cells derived from the bone marrow of wild-type mice were treated with LPS (200 ng/mL); LPS + KD, cells derived from the bone marrow of Mettl3 -KD mice were also treated with LPS (200 ng/mL). * P < 0.05, ** P < 0.01, *** P < 0.001; ns, not significant.

Article Snippet: The concentrations of IL-6, IL-17A, and tumor necrosis factor-α (TNF-α) in the supernatant of stimulated BMDCs and corneal samples were quantified using commercial mouse enzyme-linked immunosorbent assay (ELISA) kits (KE10091, KE10020, and KE10002; Proteintech, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Knockdown, Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Software, Phospho-proteomics, Derivative Assay

Journal: Molecular and cellular endocrinology

Article Title: Early transcriptome responses of the bovine midcycle corpus luteum to prostaglandin F2α includes cytokine signaling

doi: 10.1016/j.mce.2017.05.018

Figure Lengend Snippet: Predicted canonical pathways activated during the early response to PGF2α treatment.

Article Snippet: The cells were incubated in serum-free medium for 3 h before applying treatments as described in the legends to the figures [PGF2α (in ethanol, #16010, Cayman Chemical, Ann Arbor, MI), TNFα (210-TA, R&D, Minneapolis, MN), IL-1β (RP0106B), IL-6 (RP0014B), IL-17A (RP0056B, Kingfisher Biotech, Saint Paul, MN)].

Techniques: Inhibition

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 and Lipid Metabolism Correlate with Resistance to Intestinal Helminth Infection (A) C57BL/6 mice were orally infected with 200 L3 H. polygyrus ( H.p. ) larvae on day 0. A cohort of mice were sacrificed 7 days after 1° H.p. infection ( H.p. 1°). Remaining mice were drug treated (Rx) on days 14 and 15. Mice were then reinfected with H.p. on day 35 or day 56 and harvested 7 days after infection ( H.p. 2°). (B) H&E staining of the small intestine from H.p. 1° and H.p. 2° (D42). (C) Ratio-of-ratios analysis of differentially expressed genes in H.p. 1° and H.p. 2° (D42) identified distinct gene clusters (C1-3). (D) Top 10 pathways predicted to be activated more highly in and H.p. 2° than H.p. 1° (both relative to naive, 2-fold filter, p < 0.05). (E) Lipid metabolism pathway predicted activation score (relative to naive, 2-fold filter, p < 0.05). (F) Pla2g1b expression in small intestine from RNA sequencing data, confirmed by qPCR. Data are represented as mean ± SEM; n = 8, ∗ = p < 0.05, ∗∗ = p < 0.01 determined using a one-way ANOVA with Dunnett’s multiple comparison analysis. See also Figure S1 and .

Article Snippet: Recombinant mouse PLA2g1B protein , Elabscience , Cat# PKSM040953.

Techniques: Infection, Staining, Activation Assay, Expressing, RNA Sequencing, Comparison

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: Type 2 Immunity Is Insufficient for Resistance to Intestinal Helminth Infection in the Absence of Pla2g1b (A) Luminal H.p. worms in the small intestine 14 days after 2° infection. (B) Fecal egg counts 14 days after 2° H.p. infection. (C) Frequency of Il4 gfp+ CD44 hi CD4 + cells in the mesenteric lymph node (mLN) and the spleen of mice 7 days after 2° infection. (D) Frequency of cytokine + CD4 + CD44 hi cells in the mLN of mice 7 days after 2° infection. (E) ex vivo H.p. antigen-specific cytokine production from the mLN of mice 7 days after 2° infection. (F) Gene expression in the small intestine of mice 7 days after 2° H.p. infection. (G) H.p. antigen-specific IgG1 in the serum from mice 7 days after 2° H.p. infection. (H) Mucus staining (Alcian blue-periodic acid-Schiff) of the small intestine from mice 7 days after 2° H.p. infection. (I) RNA-sequencing-generated transcriptional landscape of the small intestine of WT and Pla2g1b − /− mice 7 days after 2° H.p. infection. (J) Top 10 pathways predicted to be activated 7 days after 2° H.p. infection (relative to strain-naive, 2-fold filter, p < 0.05). (K) Lipid metabolism pathway predicted activation score 7 days after 2° H.p. infection (relative to strain-naive, 2-fold filter, p < 0.05). Data are represented as mean ± SEM, n = 5–6, representative of at least three independent experiments; ns = not significant, ∗ = p < 0.05, ∗∗∗∗ = p < 0.0001 determined using a two-way ANOVA with Sidak’s multiple comparison analysis or an unpaired two-tailed t test. See also .

Article Snippet: Recombinant mouse PLA2g1B protein , Elabscience , Cat# PKSM040953.

Techniques: Infection, Ex Vivo, Gene Expression, Staining, RNA Sequencing, Generated, Activation Assay, 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: Intestinal Pla2g1b Is Regulated by the Microbiota and Rag- and Common Gamma Chain-Dependent Cells in Resistant Mice (A) Kinetics of Pla2g1b expression in the small intestine during H.p. 2° infection model. (B) Kinetics of Pla2g1b expression in the small intestine during H.p. 1° infection, n = 6. (C) Pla2g1b expression at day 28 (14 days post-Rx) in the small intestine, n = 5. (D) Pla2g1b expression at day 28 (14 days post-Rx, 1°-Rx) in the small intestine following anti-IL-4 (αIL-4) or Isotype (Iso) treatment, n = 9–10 (data pooled from two independent experiments). (E) Pla2g1b expression at day 28 (14 days post-Rx, 1° Rx) in the small intestine, n = 7–10 (data pooled from two independent experiments). (F) Pla2g1b expression at day 28 (14 days post-Rx, 1° Rx) in the small intestine following antibiotic treatment, n = 9–10 (data pooled from two independent experiments). Data are represented as mean ± SEM, n = 3. All data are representative of at least two independent experiments; ∗ = p < 0.05, ∗∗ = p < 0.01, determined using a one-way ANOVA with Dunnett’s multiple comparison analysis, unpaired t test, or a Mann-Whitney test.

Article Snippet: Recombinant mouse PLA2g1B protein , Elabscience , Cat# PKSM040953.

Techniques: Expressing, Infection, Comparison, MANN-WHITNEY

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: Epithelial-Cell-Derived Pla2g1b Is Negatively Regulated by IL-4Rα-Signaling (A) Pla2g1b detection by RNAScope ISH in the small intestine 7 days after 2° H.p. infection. (B) Pla2g1b expression in fluorescence-activated, cell sorted (FACS) CD45 – EpCam + and CD45 + EpCam − cells from the intestinal epithelium from naive and resistant (day 28) mice, n = 6. (C) Pla2g1b, Retnlb , and Gob5 expression in intestinal organoid cultures following stimulation with rIL-4 and rIL-13. Data are represented as mean ± SEM, n = 3. All data are representative of at least two independent experiments. ∗ = p < 0.05, ∗∗ = p < 0.01, ∗∗∗∗ = p < 0.0001 determined using an unpaired t test or a Mann-Whitney test. See also Figure S5 .

Article Snippet: Recombinant mouse PLA2g1B protein , Elabscience , Cat# PKSM040953.

Techniques: Derivative Assay, RNAscope, Infection, Expressing, Fluorescence, MANN-WHITNEY

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: Recombinant mouse PLA2g1B protein , Elabscience , Cat# PKSM040953.

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: Recombinant mouse PLA2g1B protein , Elabscience , Cat# PKSM040953.

Techniques: Control, Comparison, Tandem Mass Spectroscopy

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:

Article Snippet: Recombinant mouse PLA2g1B protein , Elabscience , Cat# PKSM040953.

Techniques: In Vivo, Recombinant, ATP Assay, Software, Indirect Immunoperoxidase Assay