Journal: Science translational medicine

Article Title: Steroid-induced fibroblast growth factors drive an epithelial-mesenchymal inflammatory axis in severe asthma

doi: 10.1126/scitranslmed.abl8146

Figure Lengend Snippet: (A) Normal human lung fibroblasts were cultured in 3D collagen-rich rafts, as described in Methods. Cells from five different donors were stimulated with a cocktail of FGF1, FGF2, FGF4, and FGF18, all at 10 ng/ml, for 24 hours before RNA isolation for RNA-seq. Differentially expressed genes (DEGs) where P < 0.05 are displayed. nRPKM, normalized reads per kilobase of transcript per million mapped reads. (B) Expression, in RPKM, of select DEGs is shown. *P < 0.05 by a paired t test. (C) The experimental setup depicts BAEC-ALI stimulation with F.p. (10−7 M) or diluent control for 24 hours and transfer of basal supernatant, mixed 50:50 with fresh medium, to fibroblast cultures for 24 hours. (D and E) Expression of CSF3 (D) and HAS2 (E) was measured in fibroblasts after exposure to supernatant from BAEC-ALI, as described above. Media, 100% fresh medium; Cont. Sup., 50% medium from diluent-exposed BAEC-ALI with 50% fresh medium; F.p. Sup., 50% medium from F.p.-exposed (10−7 M) BAEC-ALI with 50% fresh medium. Erdafitinib (Erd; 100 nM) was added, as indicated, to the fibroblast cultures. Data are expressed as fold change (FC) from fibroblasts cultured with medium alone. (F to H) F.p.-exposed (10−7 M) precision-cut lung slices were incubated in the presence (100 nM) or absence (diluent control) of erdafitinib. Three precision-cut lung slices were incubated per well for 24 hours, with three technical replicates per experiment. One of three experiments is shown. IL-8 (F), G-CSF (G), and hyaluronan (H) concentrations were measured by Luminex or ELISA. Error bars show SEM. *P < 0.05 with an unpaired t test.

Article Snippet: One hundred nanograms of purified RNA was reverse-transcribed using iScript (Bio-Rad) following the manufacturer’s instructions, and the following TaqMan probes (Thermo Fisher Scientific) were used: human: FGF1 , Hs01092738_m1; FGF2 , Hs00266645_m1; FGF3 , Hs00173742_m1; FGF4 , Hs00173564_m1; FGF7 , Hs00384281_m1; FGF8 , Hs00171832_m1; FGF9 , Hs00181829_m1; FGF16 , Hs00175752_m1; FGF18 , Hs00826077_m1; CSF3 , Hs99999083_m1; HAS2 , Hs00193435_m1; and FKBP5 , Hs01561003_m1; mouse: Muc5ac , Mm01276739_g1; Muc5b , Mm00466391_m1; Fkbp5 , Mm01300962_m1; Fgf1 , Mm00438906_m1; Fgf2 , Mm00433287_m1; Fgf4 , Mm00438917_m1; and Fgf18 , Mm00433286_m1.

Techniques: Cell Culture, Isolation, RNA Sequencing, Expressing, Control, Incubation, Luminex, Enzyme-linked Immunosorbent Assay

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: Cell metabolism

Article Title: Aerobic Glycolysis Controls Myeloid-Derived Suppressor Cells and Tumor Immunity via a Specific CEBPB Isoform in Triple-Negative Breast Cancer

doi: 10.1016/j.cmet.2018.04.022

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: G-CSF fragment with CMV promoter was cloned from G-CSF expression plasmid (MR225697, Origene) using KOD Xtreme hot start DNA polymerase (EMD Millipore).

Techniques: Control, Virus, Recombinant, Enzyme-linked Immunosorbent Assay, Lactate Assay, Cell Isolation, Gene Expression, shRNA, Plasmid Preparation, Software

Journal: Nucleic Acids Research

Article Title: Enhanced non-viral gene delivery by coordinated endosomal release and inhibition of β-tubulin deactylase

doi: 10.1093/nar/gkw1143

Figure Lengend Snippet: High productivity of GCSF-MSC generated with polymer based method. MSC were transfected with 200 ng GCSF DNA or 500 ng mRNA complexed with Turbofect or Lipofectamine using centrifugation method or manufacturer's instruction, respectively. Following centrifugation, the transfection mixture was replaced with media with or without DOPE/CHEMS plus 10 μM Tubastatin A. ( A ) One day later, the cells were lysed for immunoblotting analysis. ( B ) The GCSF level in the conditioned media was determined with Elisa analysis. Significant differences in GCSF level (ng/ml) were calculated using the two tailed Student's t -test. * P < 0.05. ( C ) To access the bioactivity of GCSF, the conditioned media was collected and diluted by 100 times in NSF60 culture (in serum free RPMI). After 3 day stimulation, the number of NSF60 was counted using automated cell counter (chemometec). Data presented as mean ± SD of biological triplicates. Significant differences in conditioned media from the transfected and non transfected cultures were calculated using the two tailed Student's t -test. * P < 0.05; ** P < 0.005.

Article Snippet: Cells were modified with complexes of Turbofect (ThermoScientific) with 200 ng Human Granulocyte Colony Stimulating Factor (GCSF) expression vector (SC306744, Origene) or 500 ng mRNA (Trilink).

Techniques: Generated, Transfection, Centrifugation, Western Blot, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: iScience

Article Title: Spatiotemporally organized immunomodulatory response to SARS-CoV-2 virus in primary human broncho-alveolar epithelia

doi: 10.1016/j.isci.2023.107374

Figure Lengend Snippet:

Article Snippet: Anti-CSF3 AF594 (Clone CSF3/900) , Novus , CatNBP2-47934AF594 RRID: AB_2933966.

Techniques: Bacteria, Virus, Variant Assay, Recombinant, Red Blood Cell Lysis, Software, Membrane, Cell Culture