Journal: Frontiers in Immunology

Article Title: The role of neutrophils and NETosis in lipopolysaccharide exacerbated asthmatic airway inflammation

doi: 10.3389/fimmu.2025.1651085

Figure Lengend Snippet: Deficiency in neutrophil production attenuated airway inflammation in neutrophilic asthma. (A) BALF cell counts were measured of WT mice and Csf3 -/- mice. (B) The proportion of eosinophils in two kinds of mice was determined. (C) The proportion of neutrophils was analyzed. (D) Analysis of airway resistance in two kinds of mice undergoing methacholine challenge. (E) Lung dynamic compliance was measured in each group. (F) Representative images of HE staining and PAS staining of lung tissue (scale bar=100μm). (G) Inflammatory scores of HE staining of Lung sections. (H) Percentage of PAS staining goblet cells. (I) The levels of IL6, IL8, ds-DNA, MPO-DNA in BALF. Data were shown as mean ± SD, n=6. Significance between groups was calculated using one-way ANOVA with Tukey’s post hoc method. *p<0.05, **p<0.01, ** *p<0.001 and ****p<0.0001. ns: not significant, P>0.05.

Article Snippet: Colony-stimulating factor 3 deficient ( Csf3 -/- ) and peptidyl arginine deiminase 4 deficient ( Padi4 -/- ) C57BL/6J female mice were purchased from Cyagen Biosciences (Suzhou, China).

Techniques: Staining

Journal: Advanced Science

Article Title: Reprogramming of Fatty Acid Metabolism via PPARα‐Orchestrated FADS2 in Keratinocytes Modulates Skin Inflammation in Psoriasis

doi: 10.1002/advs.202417049

Figure Lengend Snippet: PPARα acts as an upstream positive regulator of FADS2 in psoriatic keratinocytes. A) RT‐qPCR analysis of PPARA expression in normal skin from healthy controls (NN, n = 4) and lesional skin tissue from psoriasis patients (PS, n = 5). B) Representative immunofluorescence images of PPARα and K14 co‐staining in normal skin from healthy controls and lesional skin tissue from psoriasis patients. C) RT‐qPCR analysis of PPARA in psoriatic lesional skin before and 10 weeks after infliximab treatment (n = 5). D) Representative images of PPARα immunofluorescence staining in lesional skin from psoriasis patients at baseline and week 10 following infliximab treatment. E) Representative immunofluorescence images of PPARα staining in healthy skin from control mice and skin lesions from IMQ‐induced psoriasis mouse model. F,G) RT‐qPCR analysis of PPARA (F) and FADS2 (G) expression in HaCaT cells stimulated with M5 for 12 h (n = 3). H) Immunoblotting of PPARα and FADS2 in HaCaT cells stimulated with M5 cytokines for the indicated time. I) RT‐qPCR analysis of PPARA , FADS2 , and indicated genes in HaCaT cells transfected with PPARA siRNA (si PPARA ) and control siRNA (siNC) for 24 h, followed by PBS or M5 stimulation for 12 h (n = 3). J) RT‐qPCR analysis of FADS2 and indicated genes in HaCaT cells pretreated with WY14643 or DMSO for 21 h, followed by PBS or M5 stimulation for 3 h (n = 3). K) ELISA quantification of CXCL1, CXCL8, and CSF3 protein levels in cell lysates and supernatants from HaCaT cells treated as in (J) (n = 3). L) RT‐qPCR analysis of the indicated genes in HaCaT cells transfected with si FADS2 and siNC for 24 h, followed by M5 stimulation for 3 h before WY14643 or DMSO pretreatment (n = 3). Scale bar, 50 µm. Data are presented as mean ± SD. Statistical significance was determined by unpaired two‐tailed Student's t test (A,F,G,K), paired two‐tailed Student's t test (C), or one‐way ANOVA (I,J,L). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: Similarly, human CXCL1 (#70‐EK196), CSF3 (#70‐EK169), and CXCL8 (#70‐EK108/2) levels in cultured human keratinocyte lysates and supernatants were measured using corresponding human ELISA kits (Lianke Bio, China) according to the provided protocols.

Techniques: Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Control, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay, Two Tailed Test