Journal: Microbiology Spectrum

Article Title: Perilipin1 inhibits Nosema bombycis proliferation by promoting Domeless- and Hop- mediated JAK-STAT pathway activation in Bombyx mori

doi: 10.1128/spectrum.03671-23

Figure Lengend Snippet: plin1 enhances the anti- Nb effect of the JAK-STAT pathway by promoting Domeless and Hop expression. ( A, D ) Quantification of Domeless and Hop expression by RT-qPCR in BmN cells at 24, 48, and 72 hrs post- Nb infection. ( B, E ) Quantification of Domeless and Hop expression after knockdown of plin1 in BmN cells. BmN cells transfected with dsRNA-eGFP were used as a negative control. ( C, F ) Quantification of Domeless and Hop expression after overexpression of plin1 in BmN cells. BmN cells transfected with the empty vector were used as a negative control. ( G ) Quantification of Nb copy number by RT-qPCR after treatment with the JAK inhibitor at 24 and 48 h p.i. in BmN cells. BmN cells treated with DMSO were used as the negative control. ( H ) Quantification of Nb copy number by RT-qPCR after treatment with the JAK inhibitor and overexpression of plin1 in BmN cells. BmN cells treated with DMSO were used as the negative control. All the experiments were repeated three times. Error bars represent mean ± SD. One-way ANOVA or t -test, *< I>P < 0.05, **< I>P < 0.01, and ***< I>P < 0.001. ns, not significant.

Article Snippet: BmN cells were treated with 0.5–1 mM JAK inhibitor (HY-18300, Filgotinib, MCE, Nanjing, China) to inhibit JAK synthesis ( ).

Techniques: Expressing, Quantitative RT-PCR, Infection, Knockdown, Transfection, Negative Control, Over Expression, Plasmid Preparation

Journal: Cell reports

Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.

doi: 10.1016/j.celrep.2022.110420

Figure Lengend Snippet: Figure 1. Effects of filgotinib on DC responses (A) JAK1 mean fluorescence intensity (MFI) of BM-DCs stimulated with LPS over 72 h and representative histograms; n = 4. (B) JAK1 MFI of BM-DCs stimulated with the indicated TLR agonists for 24 h; n = 4 per group. (C) Cytokine profile of LPS-stimulated, filgotinib-treated BM-DCs after 24 h. Data are represented as log2 fold change compared to vehicle control; n = 4. (D) In vitro T cell proliferation assay with naive CD62L+CD44, CFSE-labeled OT-II CD4+ T cells co-cultured with vehicle- or filgotinib-treated OVA-loaded BM- DCs. Percentage of CFSElow CD4+ T cells and percentage of CD4 T cells in indicated division cycle; n = 4. (E) Representative CFSE profile of data shown in (D). (F) CD80, CD86, and MHC class II MFI of vehicle- or filgotinib-treated, LPS-stimulated BM-DCs after 24 h; n = 4. (G) Representative histograms of data shown in (F). (H) CD80, CD86, and MHC class II MFI of vehicle-, ruxolitinib-, or tofacitinib-treated, LPS-stimulated BM-DCs after 24 h; n = 4. The data are represented as means ± SEMs of biologically independent samples. **p < 0.01, ***p < 0.001, ****p < 0.0001, unpaired t test (A, D, and F), 1-way ANOVA (A and H), and 2-way ANOVA (D).

Article Snippet: Immature DCs (iDCs) were harvested on day 6. iDCs were further stimulated with LPS (100ng/ml, InvivoGen), recombinant murine IFN-ɣ (100ng/ml, #130-105-774, Miltenyi Biotech), recombinant murine IL-10 (10ng/ml, #417-ML-005, R&D), poly(I:C) LMW (1mg/ml, InvivoGen), poly(I:C) HMW (1mg/ml, InvivoGen), Imiquimod (1mg/ml, InvivoGen), CpG-A ODN 1585 (1mM, InvivoGen), CpG-B ODN 1668 (1mM, InvivoGen), Filgotinib (10mM, TargetMol), Ruxolitinib (10mM, #CAY-11609-5, Cayman Chemical), Tofacitinib (5mM #HY-40354, MedChem Express), Stattic (1mM, #HY-13818, MedChem Express), Fludarabine (100mM, #HY-B0069, MedChem Express), OVA323–399 (50mg/ml, Bachem), IFN-ɣ neutralizing antibody (1mg/ ml, Clone XMG1.2, Thermo Fisher Scientific) or isotype control antibody (1mg/ml, Thermo Fisher Scientific), InVivoMab anti-mouse IL-10R (CD210) (10mg/ml, Clone: 1B1.3A, #BE0050, BioXCell) or InVivoMAb rat IgG1 isotype control antibody (10mg/ml, Clone:HRPN, #BE0088, BioXCell).

Techniques: Control, In Vitro, Proliferation Assay, Labeling, Cell Culture

Journal: Cell reports

Article Title: JAK1 signaling in dendritic cells promotes peripheral tolerance in autoimmunity through PD-L1-mediated regulatory T cell induction.

doi: 10.1016/j.celrep.2022.110420

Figure Lengend Snippet: Figure 5. DC-expressed PD-L1 is regulated via IFN-g, JAK1, STAT1 signaling (A) PD-L1 MFI of filgotinib- or vehicle-treated or WT and JAK1-deficient BM-DCs after 24 h LPS stimulation; n = 5 per group. (B) p-STAT1(Ser727), p-STAT1(Tyr701), p-STAT3(Tyr705), PD-L1, and IRF1 kinetics following LPS stimulation of splenic WT and JAK1-deficient DCs analyzed by western blotting. (C) PD-L1 MFI of BM-DCs 24 h after IFN-ɣ stimulation in the presence of an IFN-ɣ-blocking or isotype control antibody; n = 4 per group. (D) PD-L1, p-STAT1(Ser727), and p-STAT3(Tyr705) MFI of BM-DCs after 24 h of LPS stimulation in the presence of an IFN-ɣ-blocking or isotype control antibody; n = 3–4 per group. (E) PD-L1 and p-STAT1(Ser727) MFI of WT and JAK1-deficient BM-DCs after 24 h of LPS stimulation in the presence of an IFN-ɣ-blocking or isotype control antibody; N = 3 per group. (F) PD-L1, p-STAT1(Ser727), and p-STAT3(Tyr705) MFI of BM-DCs after 24 h of LPS stimulation in the presence of a IL10-receptor-blocking or isotype control antibody; n = 3–4 per group. (G) PD-L1 MFI of BM-DCs treated with vehicle or a STAT3 inhibitor (Stattic) after 24 h of LPS stimulation; n = 4 per group. (H) PD-L1 MFI of WT and STAT1-deficient BM-DCs after 24 h of LPS stimulation. n = 4 per group. The data are represented as means ± SEMs of biologically independent samples. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, 1-way ANOVA (C), 2-way ANOVA (A and D–H).

Article Snippet: Immature DCs (iDCs) were harvested on day 6. iDCs were further stimulated with LPS (100ng/ml, InvivoGen), recombinant murine IFN-ɣ (100ng/ml, #130-105-774, Miltenyi Biotech), recombinant murine IL-10 (10ng/ml, #417-ML-005, R&D), poly(I:C) LMW (1mg/ml, InvivoGen), poly(I:C) HMW (1mg/ml, InvivoGen), Imiquimod (1mg/ml, InvivoGen), CpG-A ODN 1585 (1mM, InvivoGen), CpG-B ODN 1668 (1mM, InvivoGen), Filgotinib (10mM, TargetMol), Ruxolitinib (10mM, #CAY-11609-5, Cayman Chemical), Tofacitinib (5mM #HY-40354, MedChem Express), Stattic (1mM, #HY-13818, MedChem Express), Fludarabine (100mM, #HY-B0069, MedChem Express), OVA323–399 (50mg/ml, Bachem), IFN-ɣ neutralizing antibody (1mg/ ml, Clone XMG1.2, Thermo Fisher Scientific) or isotype control antibody (1mg/ml, Thermo Fisher Scientific), InVivoMab anti-mouse IL-10R (CD210) (10mg/ml, Clone: 1B1.3A, #BE0050, BioXCell) or InVivoMAb rat IgG1 isotype control antibody (10mg/ml, Clone:HRPN, #BE0088, BioXCell).

Techniques: Western Blot, Blocking Assay, Control

Journal: Cells

Article Title: Small Molecule Inhibitors in the Treatment of Rheumatoid Arthritis and Beyond: Latest Updates and Potential Strategy for Fighting COVID-19

doi: 10.3390/cells9081876

Figure Lengend Snippet: Kinase and other small molecule inhibitors investigated in RA.

Article Snippet: JAK1 , Filgotinib (GLPG0634, GS6034) , Galapagos NV , phase III (NCT03025308) active phase II (NCT02065700) active phase II (NCT03926195) recruiting , .

Techniques:

Journal: Scientific Reports

Article Title: IL-10 Promotes Neurite Outgrowth and Synapse Formation in Cultured Cortical Neurons after the Oxygen-Glucose Deprivation via JAK1/STAT3 Pathway

doi: 10.1038/srep30459

Figure Lengend Snippet: ( A ) Western blot analysis of p-JAK1 (n = 3). * p < 0.05, as compared with control group; # p < 0.05, as compared with OGD group; ** p < 0.01, as compared with OGD group; by one way analysis of variance (ANOVA) followed by Student-Newman-Keuls multiple comparisons test, F = 26.747, p < 0.0001. ( B ) Western blot analysis of p-STAT3 (n = 3). * p < 0.05, as compared with control group; # p < 0.05, as compared with OGD group; ## p < 0.01, as compared with OGD group; ** p < 0.01, as compared with OGD + IL-10 group; by one way analysis of variance (ANOVA) followed by Student-Newman-Keuls multiple comparisons test, F = 21.554, p < 0.0001. Data are presented as mean ± SEM.

Article Snippet: For neurons with IL-10, a specific inhibitor of JAK1, GLPG0634 (SYNkinase, USA) was added to inhibit phosphorylation of JAK1 pharmacologically, reaching a final concentration of 20 nmol/L.

Techniques: Western Blot, Control