myeloperoxidase  (Hycult Biotech)

Journal: JCI Insight

Article Title: Tamm-Horsfall protein augments neutrophil NETosis during urinary tract infection

doi: 10.1172/jci.insight.180024

Figure Lengend Snippet: Neutrophils were stained with anti–MPO-FITC, SYTOX Orange, Hoechst, and Live/Dead stain and visualized for fluorescence and bright-field (BF). ( A and B ) Gating strategy ( A ) of neutrophils subpopulations with representative images shown ( B ). ( C – G ) Frequency of NETs and NET precursors (Type III and II) ( C ), NET fragments (Type IV) ( D ), dead cells with condensed nuclei (Type V) ( E ), dead cells with decondensed nuclei (Type VI) ( F ), and live cells (Type I) ( G ). ( H ) Frequency of Hoechst + cells. ( I ) Circularity scores across groups. Experiments were performed in 8 experiments and combined, n = 8. Box-and-whisker plots show median, all points, and 25–75th percentiles ( C – I ). Data were analyzed by 1-way ANOVA with Holm-Šídák’s multiple-comparisons test ( C – I ). * P < 0.05. Sia, sialic acid.

Article Snippet: Cells were incubated with primary antibodies: sheep anti-THP polyclonal antibody (1:40, R&D Systems, AF5175) conjugated using the FITC Conjugation Kit Lightning-Link Abcam, ab102884), goat anti-MPO polyclonal antibody (1:200, R&D Systems, AF3667), and rabbit anti–Histone H3 antibody (1:100, Abcam, ab5103) in PBST+1% BSA.

Techniques: Staining, Fluorescence, Whisker Assay

Journal: Nagoya Journal of Medical Science

Article Title: Adenosine triphosphate release inhibitors targeting pannexin1 improve recovery after spinal cord injury

doi: 10.18999/nagjms.86.3.392

Figure Lengend Snippet: Immunohistochemical analysis of the injured spinal region 7 days after spinal cord injury Fig. 3A: Immunohistochemical analysis of the myeloperoxidase (MPO)-positive area in the injured spinal region 7 days after SCI. Bar: 40 µm. Fig. 3B: Quantification of Fig. 3A. The results are presented as the mean ± SEM. ** p<0.01 by Student’s t -test. Five rats were used and analyzed in each group. Fig. 3C: Immunohistochemical analysis of the CD8a-positive area in the injured spinal region 7 days after SCI. Bar: 40 µm. Fig. 3D: Quantification of Fig. 3C. The results are presented as the mean ± SEM. ** p<0.01 by Student’s t -test. Five rats were used and analyzed in each group. Fig. 3E: Immunohistochemical analysis of the CD68-positive myeloperoxidase-positive area in the injured spinal region 7 days after SCI. Bar: 40 µm. Fig. 3F: Quantification of Fig. 3E. The results are presented as the mean ± SEM. *** p<0.001 using Student’s t -test. Five rats were used and analyzed in each group. Fig. 3G: Immunohistochemical analysis of the ionized calcium-binding adaptor molecule 1 (Iba1)-positive area in the injured spinal region 7 days after SCI. Bar: 40 µm. Fig. 3H: Quantification of Fig. 3G. The results are presented as the mean ± SEM. *** p<0.001 using Student’s t -test. Five rats were used and analyzed in each group. Fig. 3I: Immunohistochemical analysis of the glial fibrillary acidic protein (GFAP)-positive area in the injured spinal region 7 days after SCI. Bar: 40 µm. Fig. 3J: Quantification of Fig. 3I. The results are presented as the mean ± SEM. *** p<0.001 using Student’s t -test. Five rats were used and analyzed in each group. BBFCF: Brilliant Blue FCF BBG: Brilliant Blue G CBX: carbenoxolone Panx1: P2X purinoceptor 7/pannexin1 SCI: spinal cord injury SEM: standard error of the mean

Article Snippet: Next, the sections were incubated with primary antibodies against ionized calcium-binding adaptor molecule 1 antibody (019-19741, 1:500, FUJIFILM, Tokyo, Japan), glial fibrillary acidic protein antibody (ab68428, 1:250, Abcam, Cambridge, UK), myeloperoxidase (HP9048-100UG, 1:50, Hycult Biotech, Uden, Netherlands), CD68 antibody (ab125212, 1:500, Abcam), and CD8a (A02236-1 1:500, Boster Bio, Pleasanton, CA, USA).

Techniques: Immunohistochemical staining, Binding Assay

Journal: Nagoya Journal of Medical Science

Article Title: Adenosine triphosphate release inhibitors targeting pannexin1 improve recovery after spinal cord injury

doi: 10.18999/nagjms.86.3.392

Figure Lengend Snippet: Immunohistochemical analysis of the injured spinal region 42 days after spinal cord injury Fig. 4A: Immunohistochemical analysis of the myeloperoxidase (MPO)-positive area in the injured spinal region 42 days after SCI. Bar: 40 µm. Fig. 4B: Quantification of Fig. 4A. The results are presented as the mean ± SEM. Twelve rats were used and analyzed in each group. Fig. 4C: Immunohistochemical analysis of the CD8a-positive area in the injured spinal region 42 days after SCI. Bar: 40 µm. Fig. 4D: Quantification of Fig. 4C. The results are presented as the mean ± SEM. Twelve rats were used and analyzed in each group. Fig. 4E: Immunohistochemical analysis of the CD68-positive area in the injured spinal region 42 days after SCI. Bar: 40 µm. Fig. 4F: Quantification of Fig. 4E. The results are presented as the mean ± SEM. Twelve rats were used and analyzed in each group. Fig. 4G: Immunohistochemical analysis of the ionized calcium-binding adaptor molecule 1 (Iba1)-positive area in the injured spinal region 42 days after SCI. Bar: 40 µm. Fig. 4H: Quantification of Fig. 4G. The results are presented as the mean ± SEM. *** p<0.001 by Student’s t -test. Twelve rats were used and analyzed in each group. Fig. 4I: Immunohistochemical analysis of the glial fibrillary acidic protein (GFAP)-positive area in the injured spinal region 42 days after SCI. Bar: 40 µm. Fig. 4J: Quantification of Fig. 4I. The results are presented as the mean ± SEM. *** p<0.001 using Student’s t -test. Twelve rats were used and analyzed in each group. BBFCF: Brilliant Blue FCF BBG: Brilliant Blue G CBX: carbenoxolone Panx1: P2X purinoceptor 7/pannexin1 N.S.: not significant according to Student’s t -test SCI: spinal cord injury SEM: standard error of the mean

Article Snippet: Next, the sections were incubated with primary antibodies against ionized calcium-binding adaptor molecule 1 antibody (019-19741, 1:500, FUJIFILM, Tokyo, Japan), glial fibrillary acidic protein antibody (ab68428, 1:250, Abcam, Cambridge, UK), myeloperoxidase (HP9048-100UG, 1:50, Hycult Biotech, Uden, Netherlands), CD68 antibody (ab125212, 1:500, Abcam), and CD8a (A02236-1 1:500, Boster Bio, Pleasanton, CA, USA).

Techniques: Immunohistochemical staining, Binding Assay