n-formyl-met-leu-phe (fmlp)  (Millipore)

Journal: Materials Today Bio

Article Title: Co-delivery of panobinostat and siSTAT3 using engineered M1 exosomes to establish a one-two punch therapeutic strategy for glioblastoma recurrence

doi: 10.1016/j.mtbio.2025.102680

Figure Lengend Snippet: Inflammation-directed targeted delivery in vitro. (A) Representative confocal images of the uptake of M1-EXOs and iM1-EXOs-PAN by GB cells. Scale bar, 25 μm. (B) Quantification of the PKH67‐positive cell ratio based on the confocal microscopy images. (C) Schematic illustration of the in vitro BBB model using a trans-well system to evaluate the penetration capability of iM1-EXOs-PAN across the endothelial monolayer. The transwell co-culture system containing HUVEC cells (simulating the vascular endothelial cells of BBB) in the upper chamber and GB cells in the bottom chamber. And, iM1-EXOs-PAN was added to the upper chamber, and FBS free medium with or without fMLP (100 μM, MCE, HY-P0224) was added to the lower chamber. (D) Representative images of HUVECs and LN229 cells up-taking the nanoformulation treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 12 h. Scale bar, 20 μm. (E) The percentage of PKH67 positive cells treated with chemotactic peptide (+fMLP, 100 μM) or none (-fMLP) at 4, 8, and 12 h. Statistical significance was calculated using two-way ANOVA. n. s = not significant, and ∗∗∗ P < 0.001.

Article Snippet: Monolayered HUVEC with TEER values greater than 300 Ω cm 2 were adopted as the BBB model. Then, 0.2 μg/μL of PKH67-labeled iM1-EXOs-PAN was added to the upper chamber, and FBS-free medium with or without fMLP (100 μΜ, HY-P0224, MCE) was added to the lower chamber [ ].

Techniques: In Vitro, Confocal Microscopy, Co-Culture Assay

Journal: International Journal of Molecular Sciences

Article Title: Neutrophils in the Spotlight—An Analysis of Neutrophil Function and Phenotype in ARDS

doi: 10.3390/ijms252312547

Figure Lengend Snippet: Overview of fluorescently labeled antibodies. Abbreviations: phycoerythrin (PE), fluorescein isothiocyanate (FITC), allophycocyanin (APC).

Article Snippet: fMLP-Receptor , APC , APC Anti-human, fMLP receptor Antibody Clone: REA169 Miltenyl Biotec, Bergisch Gladbach, Deutschland.

Techniques: Labeling

Journal: The Journal of Experimental Medicine

Article Title: Filamin A editing in myeloid cells reduces intestinal inflammation and protects from colitis

doi: 10.1084/jem.20240109

Figure Lengend Snippet: A fixed, fully FLNA R in myeloid cells shifts cellular properties and could be therapeutically exploited in colitis. (A) Flna mRNA editing status of WT whole distal colon was determined by RT-PCR amplicon Illumina sequencing on days (d) 0, 2, 5, and 10 of DSS challenge (one-way ANOVA, F = 19.09, DF = 3). (B) Gene expression analysis by real-time qPCR from WT colon homogenates. CT values relative to Gapdh (2^−ddCT) are shown as log 2 fold change compared with day zero depicted as heatmap at different time points of DSS treatment (mixed-effects analysis followed by Tukey’s multiple comparisons test). (C–F) Flna mRNA editing status of sorted cell populations of two–six pooled WT mouse colons, BM, and blood determined by RT-PCR amplicon Illumina sequencing on days 0, 2, 5, and 10 of DSS challenge (mixed-effects analysis followed by Tukey’s multiple comparisons test). (G and H) Bar graphs of cytokine levels (pg/ml) in supernatants from LPS-stimulated mouse neutrophils and BMDMs (Student’s t test). (I) Neutrophil migration speed in a pillar forest of a microfabricated PDMS device toward an fMLP gradient (Student’s t test). Dashed lines represent the median. (J) Exemplary image of CSFE- and TAMRA-stained FLNA Q and FLNA R neutrophils within the device. (K and L) Quantification of DNA release as relative fluorescence units to measure NETosis of isolated neutrophils after stimulation with a23187 or PMA over time. Time course data were plotted in a line graph with mean ± SEM (two-way ANOVA followed by Tukey’s multiple comparisons test, F = 11.94, DF = 60). (M and N) Scoring of histological colon sections stained for NETs by anti-citrullinated histone 4 (citH4) or neutrophils by anti-mouse Ly-6G. (O) Flna editing levels in intestinal tissues of WT mice: healthy mice, mice with acute colitis (day 10 of DSS), or mice recovered from colitis (day 21) were analyzed for Flna editing levels in distal and proximal colon separately ( n = 4 per time point; one-way ANOVA, F = 9.208, DF = 18). (P) FLNA editing levels in human colon biopsies of healthy donors ( n = 9), patients suffering from active UC ( n = 6), and patients in remission from active UC ( n = 6). Distal and proximal colon biopsies were analyzed separately (one-way ANOVA, distal: F = 7.326, DF = 15); comparison healthy-distal versus healthy-proximal (Student’s t test). ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05. Error bars show SEM. Data in A–F are pooled from two independent experiments ( n = 3–5 pooled colons per group, per experiment). Data in G–L are representative of two experiments, data in M and N are pooled from two experiments, and data in O are from a single experiment.

Article Snippet: N-Formyl-Met-Leu-Phe (fMLP) , Merck/MilliporeSigma , Cat#F3506.

Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Illumina Sequencing, Gene Expression, Migration, Staining, Fluorescence, Isolation, Comparison

Journal: International Journal of Molecular Sciences

Article Title: Neutrophils in the Spotlight—An Analysis of Neutrophil Function and Phenotype in ARDS

doi: 10.3390/ijms252312547

Figure Lengend Snippet: Overview of fluorescently labeled antibodies. Abbreviations: phycoerythrin (PE), fluorescein isothiocyanate (FITC), allophycocyanin (APC).

Article Snippet: fMLP-Receptor , APC , APC Anti-human, fMLP receptor Antibody Clone: REA169 Miltenyl Biotec, Bergisch Gladbach, Deutschland.

Techniques: Labeling