ll 37  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: The Alternative Polyadenylation Factor CFIm25 Orchestrates Macrophage Antibacterial Immunity by Amplifying TAB2-Mediated MAPK and NF-κB Signaling During Salmonella Infection

doi: 10.64898/2026.02.26.707986

Figure Lengend Snippet: (A) Intracellular bacterial burden (CFU) in control (−) and CFIm25-overexpressing (CFIm25-OE, +) THP-1 macrophages at two and six hours postinfection. (B) Propidium iodide flow cytometric analysis of cell death in control and CFIm25-OE macrophages at two and six hours postinfection. Graphs show the percent of cells staining with propidium iodide; only dead cells take up the stain. (C) Quantification of ROS production using the DCFH-DA fluorescence assay. (D) Quantification of NO production using the DAF-FM fluorescence assay. (E) Antimicrobial activity of conditioned media from control and CFIm25-OE macrophages against STM, assessed by CFU recovery after incubation of bacteria with media. (F) Western blot analysis of CFIm25 and LL-37 protein levels in control and CFIm25-OE macrophages at two and six hours postinfection, (G) Quantitation of LL-37 levels by densitometry measurements and normalized to GAPDH. (H) Arginase activity measured by urea production in control and CFIm25-OE macrophages at two and six hours postinfection. (I) Lactate levels in control and CFIm25-OE macrophages at two and six hours postinfection were measured using a colorimetric lactate assay kit. (J) Flow cytometric analysis of M1 (CD80) and M2 (CD206) surface marker expression in control and CFIm25-OE macrophages at two and six hours postinfection. (K) Concentrations of the TNF-α, IL-12, TGF-β, and IL-10 cytokines in culture supernatants from control and CFIm25-OE macrophages at two h and six hours postinfection as measured by ELISA. Data are presented as means ± SD from three independent experiments (n = 3); *, P < 0.05; **, P < 0.01.

Article Snippet: Antibodies used in this study included CFIm25 (Proteintech, 10322-1-AP), LL-37 (Santa Cruz, sc-166770), phosphorylated NF-κB P65 (Ser536) (Cell Signaling, 3033), NF-κB P65 (Cell Signaling, 8242), TAB2 (Cell Signaling, 3744), TBL1XR1 (Novus, NBP1-86996), phosphorylated STAT1 (Tyr701) (Cell Signaling, 9167), STAT1 (Cell Signaling, 14994), phosphorylated STAT3(Tyr705) (Cell Signaling, 9145), STAT3 (Cell Signaling, 4904), IκBα (Cell Signaling, 4814), phosphorylated P38(Thr180/Tyr182) (Cell Signaling, 4511), P38 (Cell Signaling, 9212), CD163 (Proteintech 68218-1-g), histone H3 (Cell Signaling, 9715), and GAPDH (Santa Cruz SC3233).

Techniques: Control, Staining, Fluorescence, Activity Assay, Incubation, Bacteria, Western Blot, Quantitation Assay, Lactate Assay, Marker, Expressing, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Risk Factors Associated with the Development of Thrombotic Microangiopathy in Patients with Dermatomyositis

doi: 10.3390/ijms27010315

Figure Lengend Snippet: Representative immunofluorescence confocal image of a muscle biopsy showing neutrophil extracellular traps (NETs) infiltrating the muscle tissue of dermatomyositis patient. Blue represents DNA (Column 1a , 2a , 3a ), green represents citrullinated histone H3 ( 1b , 2b , 3b ), and red represents LL37 ( 1c , 2c , 3c ). Right column images depict merged images for each tissue ( 1d , 2d , 3d ). Original magnification, ×10. Scale bar 100 µm in the first two rows and 50 µm in the third one.

Article Snippet: Then, the slides were incubated overnight at 4 °C with rabbit anti–human citrullinated histone H3 (Abcam, Cambridge, UK; dilution 1:750), and mouse anti–human LL37 (Santa Cruz Biotechnology, Dallas, TX, USA; 1: 100) diluted in 5% BSA.

Techniques: Immunofluorescence

Journal: Scientific Reports

Article Title: Proteinase 3 is involved in presepsin production through neutrophil extracellular trap phagocytosis by macrophages

doi: 10.1038/s41598-025-32574-x

Figure Lengend Snippet: Evaluation of NET formation and associated marker expression. ( a ) The percentage of the neutrophil extracellular trap (NET) area in the SS and FS plots of flow cytometry was compared among untreated neutrophils and samples following NET induction with E. coli DH5α or PMA. ( b ) Expression intensities of Cit-H3, flavocytochrome b558, SYTOX™ Green, CD14, TLR2, TLR4, and LL-37 were compared between untreated neutrophil and NET areas after E. coli DH5α stimulation. ( c ) NET ratios were compared among untreated neutrophils and samples treated with E. coli DH5α or PMA using enzyme-linked immunosorbent assay (ELISA). Data are presented as means ± standard deviations (SDs) ( n = 3). Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by Dunnett’s test. ** p < 0.01. ( d ) Western blotting was performed to examine CD14 and Cit-H3 protein levels in untreated neutrophils and those stimulated with E. coli DH5α or PMA. Band intensities were quantified using ImageJ and normalized to β-actin. Data are presented as means ± standard deviations (SDs) ( n = 3). Data are presented as means ± SDs ( n = 3). Statistical analysis was performed using one-way ANOVA followed by Tukey–Kramer’s honestly significant difference test. ** p < 0.01.

Article Snippet: Anti-human LL-37 , D-5 , – , Mouse monoclonal , sc-166,770 , Santa Cruz Biotechnology , Dallas, TX, USA.

Techniques: Marker, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot