Journal: International Journal of Biological Sciences

Article Title: DHCR24 Deficiency Drives Age-Related Meibomian Gland Dysfunction by Regulating Lipid Metabolic Imbalance and Cytosolic mtDNA-Induced cGAS-STING Activation

doi: 10.7150/ijbs.129636

Figure Lengend Snippet: Activation of cytokines and inflammatory pathways in meibocyte-specific Dhcr24 conditional knockout (cKO) mice. ( A - C ) Gene ontology (GO) analysis of differentially expressed genes (DEGs) revealing significantly enriched pathways in three categories, including ( A ) biological process (BP), ( B ) cellular component (CC), and ( C ) molecular function (MF). ( D ) Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis of DEGs between Dhcr24 fl/fl and Dhcr24 -cKO mice. ( E ) Representative immunofluorescence (IF) images of interleukin-6 (IL-6), interleukin-17 (IL-17), and tumor necrosis factor-α (TNF-α) in Dhcr24 -cKO vs. Dhcr24 fl/fl mice. Scale bars: 50 μm. ( F-H ) Enzyme-linked immunosorbent assay (ELISA) measurement of meibomian gland IL-6, IL-17, and TNF-α concentrations from Dhcr24 -cKO and Dhcr24 fl/fl mice. ( I ) Representative IF images of nuclear factor κB (NF-κB) p65 and phospho-NF-κB p65 in Dhcr24 -cKO vs. Dhcr24 fl/fl group. Scale bars: 50 μm. ( J , K ) Heatmap based on differentially expressed genes for significantly enriched pathways of ( J ) cytokines and inflammatory reponse and ( K ) NF-κB signaling. ( L ) Western blot of phospho-NF-κB p65 and NF-κB p65 protein levels. ( M ) Relative fluorescence intensity and ( N ) protein levels of phospho-NF-κB p65 and NF-κB p65.

Article Snippet: Mouse MG cryosections were permeabilized (0.1% Triton X-100), blocked with 5% BSA, and incubated with primary antibodies against KRT14 (Bioss; #bsm-52054R), DHCR24 (Bioss; #bs-5390R), IL-6 (Proteintech; 21865-1-AP), IL-17 (Bioss; #bs-1183R), TNF-α (Proteintech; #17590-1-AP), NF-κB p65 (Abcam; #ab32536), phospho-NF-kB p65 (S536) (Abcam; #ab76302), and STING (Proteintech; #19851-1-AP) overnight.

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

Journal: International Journal of Biological Sciences

Article Title: DHCR24 Deficiency Drives Age-Related Meibomian Gland Dysfunction by Regulating Lipid Metabolic Imbalance and Cytosolic mtDNA-Induced cGAS-STING Activation

doi: 10.7150/ijbs.129636

Figure Lengend Snippet: Adeno-associated virus (AAV)-mediated DHCR24 overexpression reverses ocular surface pathology in Dhcr24 conditional knockout (cKO) mice. ( A ) Experiment timeline for AAV treatment and ocular surface assessments. Two-month-old Dhcr24 -cKO and Dhcr24 fl/fl mice were injected with tamoxifen (TAM) for 7 days and raised for 2 weeks before intra-meibomian gland (MG) injection of AAV-vehicle or AAV-DHCR24, followed by a 3-month period before ocular phenotype evaluation. A separate group of two-month-old Dhcr24 -cKO mice was raised for 6 months post-TAM before undergoing the same AAV treatment and evaluation procedures. ( B ) Representative white light photos, fluorescein staining, Rose Bengal staining, and upper tarsal plate images for Dhcr24 fl/fl , young Dhcr24 -cKO (3m), and aged Dhcr24 -cKO (9m) with AAV-vehicle or AAV-DHCR24 treatment. Arrows pointing to truncated or absent MGs. ( C - F ) Ocular surface and MG scoring in the treatment groups, including ( C ) corneal opacity score ( n = 5), ( D ) corneal fluorescein staining score ( n = 5), ( E ) Rose Bengal staining score ( n = 5), and ( F ) clinical MG score ( n = 5). ( G ) Oil Red O staining of upper tarsal plate sections in the different treatment groups, with circles indicating absent MGs. Scale bars: 200 μm. ( H ) Hematoxylin-eosin (H&E) staining comparing the Dhcr24 -cKO mice treated with AAV-vehicle and AAV-DHCR24. Scale bars: 50 μm. ( I ) Immunofluorescence (IF) staining and ( J ) relative fluorescence intensity of STING for young Dhcr24 -cKO (3m) and aged Dhcr24 -cKO (9m) treated with either AAV-vehicle or AAV-DHCR24. Scale bars: 50 μm. ( K-M ) Relative mRNA expression of ( K ) interleukin-6 (IL-6), ( L ) interleukin-17 (IL-17), and ( M ) tumor necrosis factor-α (TNF-α) in mouse MGs of Dhcr24 -cKO (3m) and Dhcr24 -cKO (9m) following AAV-vehicle or AAV-DHCR24 treatment.

Article Snippet: Mouse MG cryosections were permeabilized (0.1% Triton X-100), blocked with 5% BSA, and incubated with primary antibodies against KRT14 (Bioss; #bsm-52054R), DHCR24 (Bioss; #bs-5390R), IL-6 (Proteintech; 21865-1-AP), IL-17 (Bioss; #bs-1183R), TNF-α (Proteintech; #17590-1-AP), NF-κB p65 (Abcam; #ab32536), phospho-NF-kB p65 (S536) (Abcam; #ab76302), and STING (Proteintech; #19851-1-AP) overnight.

Techniques: Virus, Over Expression, Knock-Out, Injection, Staining, Immunofluorescence, Fluorescence, Expressing

Journal: Frontiers in Immunology

Article Title: Intramuscular Priming and Intranasal Boosting Induce Strong Genital Immunity Through Secretory IgA in Minipigs Infected with Chlamydia trachomatis

doi: 10.3389/fimmu.2015.00628

Figure Lengend Snippet: Lymph node IFN-γ and IL-17A response to Hirep1 re-stimulation at day 15 pi .

Article Snippet: IL-17A was detected with a rabbit anti-pig IL-17A polyclonal-coating antibody (KP0498S-100, Kingfischer Biotech, Saint Paul) (1 μg/ml) biotinylated rabbit anti-pig IL-17A polyclonal detection antibody (KPB0499S-050, Kingfisher Biotech, Saint Paul) in 0.1 μg/ml and HRP-conjugated streptavidin (SNN1004, Life TechnologiesTM, Denmark) in 0.1 μg/ml.

Techniques:

Journal: Frontiers in Immunology

Article Title: Intramuscular Priming and Intranasal Boosting Induce Strong Genital Immunity Through Secretory IgA in Minipigs Infected with Chlamydia trachomatis

doi: 10.3389/fimmu.2015.00628

Figure Lengend Snippet: Lymph node IFN-γ and IL-17A response to Hirep1 re-stimulation at day 15 pi .

Article Snippet: IL-17A was detected with a rabbit anti-pig IL-17A polyclonal-coating antibody (KP0498S-100, Kingfischer Biotech, Saint Paul) (1 μg/ml) biotinylated rabbit anti-pig IL-17A polyclonal detection antibody (KPB0499S-050, Kingfisher Biotech, Saint Paul) in 0.1 μg/ml and HRP-conjugated streptavidin (SNN1004, Life TechnologiesTM, Denmark) in 0.1 μg/ml.

Techniques:

Journal: Journal of Translational Medicine

Article Title: The treatment of Qibai Pingfei Capsule on chronic obstructive pulmonary disease may be mediated by Th17/Treg balance and gut-lung axis microbiota

doi: 10.1186/s12967-022-03481-w

Figure Lengend Snippet: Cytokines and transcription factors related to Th17 cells and Treg cells. ELISA showing IL-17A, IL-10, CCL20, and CCR6 protein expression levels ( A ). Western blot analysis and quantification of western blotting showing RORγt and Foxp3 ( B ). The immunohistochemistry staining and the mean optical density of RORγt ( C ) and Foxp3 ( D ) in the lung tissue. Data are expressed as mean ± standard deviation (SD). * P < 0.05, ** P < 0.01

Article Snippet: Then use fluorescein isothiocyanate (FITC)-conjugated anti-CD4 (cat.no.201505; clone.no.W3/25; Biolegend) as the abscissa and phycoerythrin (PE)-conjugated anti-interleukin (IL)-17A (cat.no.bs-1183R-PE; Bioss) as the ordinate to mark the CD4 + IL-17 + Th17 cells.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Immunohistochemistry, Staining, Standard Deviation