alexa fluor 594 anti mouse s100a9  (Novus Biologicals)

Journal: Biomarker Research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Screening and analysis of the core target molecule S100A9. ( A ) Venny plot of the top 10 genes upregulated in the EAC group and 18 candidate key genes for IC/BPS. ( B - C ) Analysis of S100A9 expression in the bladders from EAC mice and IC/BPS patients. ( D ) ROC curve analysis of S100A9 gene expression in the ulcer and non-ulcer groups of patients with IC/BPS. ( E ) Immunohistochemical analysis of S100A9 expression in EAC mice (×200, n = 6). ( F ) Western blot analysis of S100A9 expression in EAC mice. ( G ) Immunohistochemical analysis of S100A9 expression in IC/BPS patients (×200, n = 6). NS indicates no difference; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The antibodies used in this investigation were as follows: rabbit monoclonal anti-S100A9 antibody (1:400, 73425, CST, USA), mouse monoclonal anti-CD4 antibody (1:100, Thermo Fisher, USA), mouse monoclonal anti-68 antibody (1:100, sc-20060, Santa Cruz Biotechnology, USA), rabbit anti-F4/80 antibody (1:100, ab300421, abcam, USA), rabbit polyclonal anti-S100A9 antibody (1:600, 26992-1-AP, Proteintech, China) and Universal Reagent Kit mouse mouse/rabbit polymer detection system (PV-6000, Zhongshan Inc, China).

Techniques: Expressing, Gene Expression, Immunohistochemical staining, Western Blot

Journal: Biomarker Research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Expression analysis of S100A9 in EAC group from data of single-cell sequencing. ( A ) Venny plot of overlapping genes related to S100A9 and S100A8 comparing within the EAC and control groups. ( B ) GO enrichment analysis of gene pathways associated with S100A9 and S100A8. ( C ) Distribution and expression analysis of S100A9 and S100A8 in different bladder cell types between control and EAC group. ( D ) Comparative analysis of S100A9 and S100A8 expression in macrophages between control and EAC groups. ( E ) Expression analysis of TLR4 receptor in bladder cells between control and EAC groups. ( F ) Comparative analysis of gene expression in macrophages between the EAC and control groups. ( G ) GSEA analysis of the activation pathway of MACROPHAGE ACTIVATION. ( H ) Analysis of cell-cell communication source of neutrophils between control and EAC group. ( I ) Analysis of cell-cell communication source of macrophages between control and EAC group. ( J ) Immunofluorescence confocal analysis of S100A9 and macrophage marker F4/80 in control and EAC groups ( n = 6)

Article Snippet: The antibodies used in this investigation were as follows: rabbit monoclonal anti-S100A9 antibody (1:400, 73425, CST, USA), mouse monoclonal anti-CD4 antibody (1:100, Thermo Fisher, USA), mouse monoclonal anti-68 antibody (1:100, sc-20060, Santa Cruz Biotechnology, USA), rabbit anti-F4/80 antibody (1:100, ab300421, abcam, USA), rabbit polyclonal anti-S100A9 antibody (1:600, 26992-1-AP, Proteintech, China) and Universal Reagent Kit mouse mouse/rabbit polymer detection system (PV-6000, Zhongshan Inc, China).

Techniques: Expressing, Sequencing, Control, Gene Expression, Activation Assay, Immunofluorescence, Marker

Journal: Biomarker Research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Analysis of S100A9 expression and immune cell infiltration in IC/BPS bladder specimens. Analysis of HE staining of bladder in patients with ulcer type of IC/BPS (×200, n = 6). ( B ) Analysis of bladder mast cell staining in patients with ulcer type of IC/BPS (×200, n = 6). ( C ) Analysis of CD68 staining of bladder macrophages from ulcer type IC/BPS patients (×200, n = 6). ( D ) CD4 staining analysis of bladder T cells from patients with ulcer type of IC/BPS (×200, n = 6). ( E ) Confocal immunofluorescence analysis of S100A9 and CD68 in normal and IC/BPS groups ( n = 6). NS indicates no difference; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The antibodies used in this investigation were as follows: rabbit monoclonal anti-S100A9 antibody (1:400, 73425, CST, USA), mouse monoclonal anti-CD4 antibody (1:100, Thermo Fisher, USA), mouse monoclonal anti-68 antibody (1:100, sc-20060, Santa Cruz Biotechnology, USA), rabbit anti-F4/80 antibody (1:100, ab300421, abcam, USA), rabbit polyclonal anti-S100A9 antibody (1:600, 26992-1-AP, Proteintech, China) and Universal Reagent Kit mouse mouse/rabbit polymer detection system (PV-6000, Zhongshan Inc, China).

Techniques: Expressing, Staining, Immunofluorescence

Journal: Biomarker Research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: S100A9 promotes macrophage activation and secretion of inflammatory factors. ( A ) LPS promoted S100A9 secretion in mice primary macrophages ( n = 6). ( B - C ) S100A9 promoted pro-inflammatory polarisation of primary mouse macrophages ( n = 3). ( D ) S100A9 promoted IL-6, TNF-α, and IL-1β expression in mouse primary macrophages ( n = 6). ( E ) Paquinimod inhibited the increased IL-6, TNF-α and IL-1β mRNA expression in S100A9-induced mouse macrophages ( n = 9). ( F ) Paquinimod inhibited the increase of IL-6, TNF-α and IL-1β levels in S100A9-induced mouse macrophages ( n = 9). ( G ) S100A9 activated TLR4/NF-κB and TLR4/p38 signalling pathways in primary mouse macrophages ( n = 3). NS indicates no difference; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The antibodies used in this investigation were as follows: rabbit monoclonal anti-S100A9 antibody (1:400, 73425, CST, USA), mouse monoclonal anti-CD4 antibody (1:100, Thermo Fisher, USA), mouse monoclonal anti-68 antibody (1:100, sc-20060, Santa Cruz Biotechnology, USA), rabbit anti-F4/80 antibody (1:100, ab300421, abcam, USA), rabbit polyclonal anti-S100A9 antibody (1:600, 26992-1-AP, Proteintech, China) and Universal Reagent Kit mouse mouse/rabbit polymer detection system (PV-6000, Zhongshan Inc, China).

Techniques: Activation Assay, Expressing

Journal: Biomarker Research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Analysis of the effects of S100A9 knockdown on bladder tissue and immune cells in EAC mice ( n = 6). (A) HE staining analysis of bladder tissues in WT and S100A9 knockdout mice of control and EAC groups (×200). ( B ) Analysis of mast cell infiltration in WT and S100A9 knockout bladder tissues from control and EAC groups (×200). ( C ) TUNEL staining for apoptosis analysis in bladder tissues from WT and S100A9 knockout mice in the control and EAC groups (×200). ( D ) Immunohistochemical analysis of macrophage marker F4/80 in WT and S100A9 knockout mice of control and EAC groups (×200). ( E ) Immunohistochemical analysis of CD4 in WT and S100A9 knockout mice of control and EAC groups (×200). ( F ) Immunohistochemical analysis of S100A9 in WT and S100A9 knockdout mice of the control and EAC groups (×200). NS indicates no difference; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The antibodies used in this investigation were as follows: rabbit monoclonal anti-S100A9 antibody (1:400, 73425, CST, USA), mouse monoclonal anti-CD4 antibody (1:100, Thermo Fisher, USA), mouse monoclonal anti-68 antibody (1:100, sc-20060, Santa Cruz Biotechnology, USA), rabbit anti-F4/80 antibody (1:100, ab300421, abcam, USA), rabbit polyclonal anti-S100A9 antibody (1:600, 26992-1-AP, Proteintech, China) and Universal Reagent Kit mouse mouse/rabbit polymer detection system (PV-6000, Zhongshan Inc, China).

Techniques: Knockdown, Staining, Control, Knock-Out, TUNEL Assay, Immunohistochemical staining, Marker

Journal: Biomarker Research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Knockdown of S100A9 significantly reduced TLR4/NF-κB and TLR4/p38 signalling pathway activating and decreasing inflammation and apoptosis-related protein expression in EAC mice ( n = 6). GSEA analysis showing that TLR4/MyD88, NF-κB, and p38 signalling are activated in IC/BPS patients and EAC mice. ( B - C ) Knockdown of S100A9 significantly reduced TLR4/NF-κB and TLR4/p38 signalling pathway protein expression in EAC mice. ( D - E ) Knockdown of S100A9 significantly reduced the expression of bladder inflammation-related proteins (IL-6, IL-1β and TNF-α) in EAC mice. ( F ) GSEA analysis showing that apoptosis signalling is activated in IC/BPS patients and EAC mice. ( G - H ) Knockdown of S100A9 expression significantly reduced the expression of bladder apoptosis-related proteins (Bax, caspase-3, caspase-8, and caspase-1) and improved the expression of epithelial damage marker proteins (UPK3A and UPK2) in EAC mice. NS indicates no difference; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The antibodies used in this investigation were as follows: rabbit monoclonal anti-S100A9 antibody (1:400, 73425, CST, USA), mouse monoclonal anti-CD4 antibody (1:100, Thermo Fisher, USA), mouse monoclonal anti-68 antibody (1:100, sc-20060, Santa Cruz Biotechnology, USA), rabbit anti-F4/80 antibody (1:100, ab300421, abcam, USA), rabbit polyclonal anti-S100A9 antibody (1:600, 26992-1-AP, Proteintech, China) and Universal Reagent Kit mouse mouse/rabbit polymer detection system (PV-6000, Zhongshan Inc, China).

Techniques: Knockdown, Expressing, Marker

Journal: Biomarker Research

Article Title: S100A9 as a potential novel target for experimental autoimmune cystitis and interstitial cystitis/bladder pain syndrome

doi: 10.1186/s40364-025-00763-5

Figure Lengend Snippet: Paquinimod inhibition and S100A9 knockdown significantly improved bladder function in EAC mice ( n = 6). ( A ) Analysis of cystometry data from control, EAC and paquinimod treatment groups. ( B ) Comparative analysis of micturition frequency (MF), maximum bladder pressure (MBP) and inter-contraction interval (ICI) in the control, EAC and paquinimod treatment groups. ( C ) Analysis of cystometry data of WT, S100A9 −/− , EAC and S100A9 −/− +EAC groups. ( D ) Comparative analysis of MF, MP and ICI in the WT, S100A9 −/− , EAC and paquinimod treatment groups. NS indicates no difference; * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: The antibodies used in this investigation were as follows: rabbit monoclonal anti-S100A9 antibody (1:400, 73425, CST, USA), mouse monoclonal anti-CD4 antibody (1:100, Thermo Fisher, USA), mouse monoclonal anti-68 antibody (1:100, sc-20060, Santa Cruz Biotechnology, USA), rabbit anti-F4/80 antibody (1:100, ab300421, abcam, USA), rabbit polyclonal anti-S100A9 antibody (1:600, 26992-1-AP, Proteintech, China) and Universal Reagent Kit mouse mouse/rabbit polymer detection system (PV-6000, Zhongshan Inc, China).

Techniques: Inhibition, Knockdown, Control