Journal: Journal of Translational Medicine

Article Title: Tumor-associated macrophage-specific LRRFIP1 promotes M2 macrophage polarization and progression of colorectal cancer via activation of the PI3K pathway

doi: 10.1186/s12967-026-07759-1

Figure Lengend Snippet: High expression of LRRFIP1 in colorectal cancer invasive front and ex vivo co-cultured tumor-associated macrophages (TAMs). ( A ) Violin plot of LRRFIP1 expression in macrophages of different tissues. ( B ) Representative immunofluorescence staining of LRRFIP1 expression in patient colorectal cancer and adjacent normal tissue. Red, CD163; green, LRRFIP1; blue, DAPI. Scale bar: 500 μm, 100 μm. ( C ) Statistical analysis of LRRFIP1 + /CD163 + TAM ratio in different tissues, n = 5. ( D ) Survival probability between LRRFIP1 high and LRRFIP1 low patients. ( E - F ) Flow cytometry analysis of CD163 in different treatment groups of THP-1. The detailed gating strategy is shown in (Fig. A). ( G ) LRRFIP1 mRNA expression levels in the THP-1 of different treatment groups were analyzed by real-time quantitative PCR and normalized to β-actin. ( H - I ) The protein levels of LRRFIP1 and β-actin in THP-1 were determined by WB. β-actin was used as a sample control. The data for each group came from three independent measurements and are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, ns: no significant

Article Snippet: Anti-mTOR (AF6308), Anti-Phospho-mTOR (Ser2448) (AF3308), Anti-AKT (AF0836), Anti-Phospho-AKT(Ser473) (AF0016), anti-IL-10 (DF6894), anti-E-cadherin (AF0131), anti-MMP9 (AF5228), anti-BAX(AF0120), anti-Ki67(AF0198), anti-Vimentin (AF7013), anti-CD163 Antibody (DF8235), Goat Anti-Rabbit IgG (H + L) Fluor594-conjugated (S0006) and Goat Anti-Mouse IgG (H + L) Fluor488-conjugated (S0017) were purchased from Affinity Biosciences, anti-LRRFIP1(sc-515571) was purchased from Santa Cruz biotechnology, anti-SUCNR1/GPR91 Antibody (NBP1-00861) were purchased from Novus Biologicals. was purchased from Thermo Fisher Scientific.

Techniques: Expressing, Ex Vivo, Cell Culture, Immunofluorescence, Staining, Flow Cytometry, Real-time Polymerase Chain Reaction, Control

Journal: Journal of Translational Medicine

Article Title: Tumor-associated macrophage-specific LRRFIP1 promotes M2 macrophage polarization and progression of colorectal cancer via activation of the PI3K pathway

doi: 10.1186/s12967-026-07759-1

Figure Lengend Snippet: Interference with LRRFIP1 in ex vivo co-cultured TAMs reduces M2 phenotype and inhibits the PI3K pathway. ( A - B ) LRRFIP1 and IL-10 mRNA expression levels in THP-1 of different treatment groups were analyzed by real-time quantitative PCR and normalized to β-actin. ( C ) The protein levels of LRRFIP1, m-TOR, p-mTOR, AKT, p-AKT, IL-10 and β-actin in THP-1 were determined by WB. β-actin was used as a sample control. ( D - E ) Flow cytometry analysis of CD163 in THP-1 - derived macrophages, which were generated by co-culturing with HCT116 cells, under different treatment conditions. ( F ) ELISA assay for TGF-β level in the supernatant of macrophage, and si-LRRFIP1 macrophage with or without cocultured with HCT116, error bars indicate SEM per group in one experiment. The data for each group came from three independent measurements are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Anti-mTOR (AF6308), Anti-Phospho-mTOR (Ser2448) (AF3308), Anti-AKT (AF0836), Anti-Phospho-AKT(Ser473) (AF0016), anti-IL-10 (DF6894), anti-E-cadherin (AF0131), anti-MMP9 (AF5228), anti-BAX(AF0120), anti-Ki67(AF0198), anti-Vimentin (AF7013), anti-CD163 Antibody (DF8235), Goat Anti-Rabbit IgG (H + L) Fluor594-conjugated (S0006) and Goat Anti-Mouse IgG (H + L) Fluor488-conjugated (S0017) were purchased from Affinity Biosciences, anti-LRRFIP1(sc-515571) was purchased from Santa Cruz biotechnology, anti-SUCNR1/GPR91 Antibody (NBP1-00861) were purchased from Novus Biologicals. was purchased from Thermo Fisher Scientific.

Techniques: Ex Vivo, Cell Culture, Expressing, Real-time Polymerase Chain Reaction, Control, Flow Cytometry, Derivative Assay, Generated, Enzyme-linked Immunosorbent Assay

Journal: Journal of Translational Medicine

Article Title: Tumor-associated macrophage-specific LRRFIP1 promotes M2 macrophage polarization and progression of colorectal cancer via activation of the PI3K pathway

doi: 10.1186/s12967-026-07759-1

Figure Lengend Snippet: Blocking the PI3K/AKT pathway negates the selective activation of macrophages by LRRFIP1 and TGF-β secretion. ( A ) IL-10 mRNA expression levels in si-LRRFIP1 cells under LY294002 treatment. ( B ) IL-10 and PI3K pathway associated protein AKT, p-AKT, m-TOR, p-mTOR levels were measured by WB in different treatment groups of cells cocultured with HCT116. ( C - D ) Flow cytometry analysis of CD163 in different treatment groups of TAMs. ( E ) ELISA assay for TGF-β level in the supernatant of NC and LRRFIP1-interference TAM pretreated with DMSO or LY294002. The data for each group came from three independent measurements and are presented as mean ± SD. ** p < 0.01, *** p < 0.001, ns: no significant

Article Snippet: Anti-mTOR (AF6308), Anti-Phospho-mTOR (Ser2448) (AF3308), Anti-AKT (AF0836), Anti-Phospho-AKT(Ser473) (AF0016), anti-IL-10 (DF6894), anti-E-cadherin (AF0131), anti-MMP9 (AF5228), anti-BAX(AF0120), anti-Ki67(AF0198), anti-Vimentin (AF7013), anti-CD163 Antibody (DF8235), Goat Anti-Rabbit IgG (H + L) Fluor594-conjugated (S0006) and Goat Anti-Mouse IgG (H + L) Fluor488-conjugated (S0017) were purchased from Affinity Biosciences, anti-LRRFIP1(sc-515571) was purchased from Santa Cruz biotechnology, anti-SUCNR1/GPR91 Antibody (NBP1-00861) were purchased from Novus Biologicals. was purchased from Thermo Fisher Scientific.

Techniques: Blocking Assay, Activation Assay, Expressing, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Immunology

Article Title: Identification and validation of immune and diagnostic biomarkers for interstitial cystitis/painful bladder syndrome by integrating bioinformatics and machine-learning

doi: 10.3389/fimmu.2025.1511529

Figure Lengend Snippet: Hub genes for pediatric sepsis diagnosis in IC/BPS patients. (A) Venn diagram showed the intersection of diagnostic markers obtained by the three algorithms. (B) Nomogram is used to predict the occurrence of IC/BPS. (C) DCA curves of the diagnostic efficacy verification; (D) Calibration curve of the diagnostic efficacy verification; (E) Clinical impact curve of the diagnostic efficacy verification. (F) The ROC curve of PLAC8 in IC/BPS dataset; (G) The ROC curve of PPBP in IC/BPS dataset. (H) The ROC curve of S100A8 in IC/BPS dataset. (I) Boxplot showed the expression of PLAC8 between IC/BPS group and control group. (J) Boxplot showed the expression of PPBP between IC/BPS group and control group. (K) Boxplot showed the expression of S100A8 between IC/BPS group and control group. IC/BPS, interstitial cystitis/painful bladder syndrome; DCA, decision curve analysis; CIC, clinical impact curve; ROC, receiver operating characteristic. ***, P < 0.001.

Article Snippet: The primary antibodies utilized in this study included PLAC8 (cat. 12284-1-AP, Proteintech, China), CXCL10 (cat. DF6417, Affinity, USA), c-Kit (CD117) (cat. AF6153, Affinity, USA), SDC1 (CD138) (cat. DF6367, Affinity, USA), CD163 (cat. DF8235, Affinity, USA), CD20 (cat. DF13319, Affinity, USA), CD14 (cat. DF13278, Affinity, USA), FucT-IV (CD15) (cat. DF8547, Affinity, USA), and CD3D (cat. DF6370, Affinity, USA).

Techniques: Biomarker Discovery, Diagnostic Assay, Expressing, Control

Journal: Frontiers in Immunology

Article Title: Identification and validation of immune and diagnostic biomarkers for interstitial cystitis/painful bladder syndrome by integrating bioinformatics and machine-learning

doi: 10.3389/fimmu.2025.1511529

Figure Lengend Snippet: Verification of PLAC8 expression and immune cell infiltration in verified cohort. (A) Representative results of protein expression levels of PLAC8, CXCL10 and 7 immune cell markers in IC/BPS tissue and UC tissue (Microscale: 100 μm). (B) The IHC score of PLAC8 in IC/BPS tissues and UC tissues. (C) The IHC score of CXCL10 in IC/BPS tissues and UC tissues. (D) Number of positive cells for seven immune cell marker in IC/BPS tissues and UC tissues. (E) The relative proportions of seven immune cell marker’s protein expression in IC/BPS tissues and UC tissues. IC/BPS, interstitial cystitis/painful bladder syndrome; UC, unaffected control; IHC, Immunohistochemistry. **P < 0.01; ***P < 0.001.

Article Snippet: The primary antibodies utilized in this study included PLAC8 (cat. 12284-1-AP, Proteintech, China), CXCL10 (cat. DF6417, Affinity, USA), c-Kit (CD117) (cat. AF6153, Affinity, USA), SDC1 (CD138) (cat. DF6367, Affinity, USA), CD163 (cat. DF8235, Affinity, USA), CD20 (cat. DF13319, Affinity, USA), CD14 (cat. DF13278, Affinity, USA), FucT-IV (CD15) (cat. DF8547, Affinity, USA), and CD3D (cat. DF6370, Affinity, USA).

Techniques: Expressing, Marker, Control, Immunohistochemistry

Journal: Frontiers in Immunology

Article Title: Identification and validation of immune and diagnostic biomarkers for interstitial cystitis/painful bladder syndrome by integrating bioinformatics and machine-learning

doi: 10.3389/fimmu.2025.1511529

Figure Lengend Snippet: The expression and biological functions of PLAC8 in IC/BPS were further validated through in vitro experiments. (A) Representative results of the expression level of PLAC8 protein in inflammatory cell model was detected by western blot. (B) Representative results of PLAC8 overexpression in SV-HUC-1 cells by western blot. (C) Representative results of AKT/mTOR/PI3K pathway marker proteins were detected by western blot in SV-HUC-1 cells. (D) Proliferation of SV-HUC-1 cells was detected by cell counting kit-8. (E) Representative results of migration ability were measured by wound healing assay in SV-HUC-1 cells (Microscale: 500 μm). OD, optical density; NC, negative control; OE, over expression. **P < 0.01; ##P < 0.01; ###P < 0.001.

Article Snippet: The primary antibodies utilized in this study included PLAC8 (cat. 12284-1-AP, Proteintech, China), CXCL10 (cat. DF6417, Affinity, USA), c-Kit (CD117) (cat. AF6153, Affinity, USA), SDC1 (CD138) (cat. DF6367, Affinity, USA), CD163 (cat. DF8235, Affinity, USA), CD20 (cat. DF13319, Affinity, USA), CD14 (cat. DF13278, Affinity, USA), FucT-IV (CD15) (cat. DF8547, Affinity, USA), and CD3D (cat. DF6370, Affinity, USA).

Techniques: Expressing, In Vitro, Western Blot, Over Expression, Marker, Cell Counting, Migration, Wound Healing Assay, Negative Control

Journal: Journal of the National Cancer Center

Article Title: S100A8 promotes tumor progression by inducing phenotypic polarization of microglia through the TLR4/IL-10 signaling pathway in glioma

doi: 10.1016/j.jncc.2024.07.001

Figure Lengend Snippet: S100A8 binds TLR4 to promote IL-10 secretion. (A) Cytoscape was used to build the protein-protein interaction network of co-expressed genes. (B) Genecards was used to explore genes related to S100A8 and macrophage polarization. (C) Western blot was used to detect the expression levels of TLR4 and IL-10 after S100A8 knockdown and overexpression. (D) The content of IL-10 in cell supernatant was detected by ELISA. (E, F) Verify the combined ability of the S100A8 and TLR4 using coimmunoprecipitation and immunocytochemistry. Scale bar, 10 µm. (G, H) After adding TLR4 inhibitor TLR-IN—C34, the content of IL-10 in and out of cells was detected by Western blot and ELISA. (I) After treatment with TLR4 inhibitors, glioma cells were co-cultured with microglia, and the mRNA expression levels of M1 and M2 markers were detected by qPCR. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: The membranes were incubated in sealing fluid for 10 min at room temperature and then with primary antibodies against S100A8 (Cat. #ab288715; Abcam; 1:1000), CD163 (Cat. #DF8235; Affinity; 1:1000), ARG1 (Cat. #DF6657; Affinity; 1:1000), TLR4 (Cat. #66350-1-Ig; Proteintech; 1:5000), IL-10 (Cat. #60269-1-Ig; Proteintech; 1:5000), p65 (Cat. #80979-7-RR; Proteintech; 1:5000), p-p65 (Cat. #3033S; Cell Signaling Technology; 1:1000), JAK1 (Cat. #AF5012; Affinity; 1:1000), STAT3 (Cat. #AF6294; Affinity; 1:1000), and β-actin (Cat. #81115-1-RR; Proteintech; 1:5000) at 4°C overnight.

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Enzyme-linked Immunosorbent Assay, Immunocytochemistry, Cell Culture