e. coli serotype o11  (InvivoGen)

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Spatial heterogeneity of immune cells in the tumor microenvironment. Immunophenotypic profiling across invasion states: Non-invaded pseudocapsule (n=9; pathologically tumor-free), tumor-invaded pseudocapsule (n=23), control dura mater from non-invasive tumor cases (n=10), tumor-invaded dura mater (n=21), tumor-invaded mucosa (n=10) and non-invaded mucosa (n=53). (A) Representative immunohistochemistry images for the detection of macrophages (IBA-1 + ), CD4 + T cells, CD8 + T cells and CD19 + B cells. Arrows indicate CD19-positive cells. (B) Quantification of macrophage burden (IBA-1 + immunoreactive area; %). (C) Quantification of CD4 + T-cell density (cells per HPF). (D) Quantification of CD8 + T-cell density (cells per HPF). (E-J) Spatial heterogeneity of macrophage phenotypes. (E) Multiplex immunofluorescence images showing IBA-1 + (red), HLA-DR + (green; M1-like) and CD206 + (magenta; M2-like) macrophage distributions at the IF of the pseudocapsule, dura mater and mucosa, and in non-invaded mucosa. (F) Grayscale-intensity distributions for IBA-1 quantified using ImageJ. (G) M1 immunoreactive area (% of microscopic field) in each group (TIM, TIM-IF, TIDM, TIDM-IF, TIP and TIP-IF). (H) M2 immunoreactive area (% of microscopic field) in each group (TIM, TIM-IF, TIDM, TIDM-IF, TIP and TIP-IF). (I) M1 immunoreactive area (% of microscopic field) in the TIM and NIM groups. (J) M2 immunoreactive area (% of microscopic field) in the TIM and NIM groups. (B-D) Kruskal-Wallis test with prespecified Dunn's post hoc planned comparisons (NIM vs. TIM/NIP/TIP/NIDM/TIDM) and Bonferroni correction. (G and H) Unpaired comparisons among TIM, TIDM and TIP, and separately among TIM-IF, TIDM-IF and TIP-IF, were performed using the Kruskal-Wallis test followed by Dunn's multiple-comparisons test, whereas paired comparisons between each tumor region and its matched IF region were performed using the two-tailed Wilcoxon signed-rank test. Bonferroni correction was applied across all nine comparisons performed in this analysis. (I and J) Unpaired comparisons were analyzed using a two-tailed Mann-Whitney U test. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001. HPF, high-power field; IBA-1, ionised calcium binding adaptor molecule 1; IF, invasive front; NIDM, non-invaded dura mater; NIM, non-invaded mucosa; NIP, non-invaded pseudocapsule; ns, not significant; TIDM, tumor-invaded dura mater; TIM, tumor-invaded mucosa; TIP, tumor-invaded pseudocapsule.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Control, Immunohistochemistry, Multiplex Assay, Immunofluorescence, Two Tailed Test, MANN-WHITNEY, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Elevated IgG levels drive macrophage M2-to-M1 reprogramming. (A) Sphenoid sinus-invasive tumor cases stratified into CD19-high (n=5) and CD19-low (n=5) groups based on the cohort median of CD19 + B cell density, with (B) quantitative analyses of macrophage polarization (M1-like versus M2-like). (C) Dural-invasive tumor and non-invasive tumor cases stratified into IgG-high (n=27) and IgG-low (n=26) groups based on the cohort median of relative IgG immunohistochemistry staining intensity, with (D) quantitative analyses of M1-like/M2-like macrophage proportions. (E and F) RAW264.7 macrophages were pre-polarized with IL-4 (20 ng/ml) or with lipopolysaccharide (100 ng/ml) plus IFN-γ (20 ng/ml) for 24 h, followed by IgG (10 µg/ml) exposure. Relative (E) IL-6 and (F) TNF-α mRNA expression in RAW264.7 macrophages pre-polarized to M0, M1 or M2 states. (G) Representative flow cytometric cell-cycle profiles of TtT/GF cells following the indicated treatments. (H) Stacked bar plot summarizing the percentages of cells from (G) in G 1 , S and G 2 /M phases. (I) Representative images from the scratch wound assay at 0, 24, 48 and 72 h under the indicated treatments. (J) Quantification of scratch wound closure. (K) Representative western blot images showing total STAT1, p-STAT1, total STAT3, p-STAT3 and β-actin levels in cells treated with IFN-γ (100 ng/ml), IL-6 (100 ng/ml), IFN-γ + IL-6 (50 ng/ml each), ruxolitinib (5 µM) or IFN-γ + IL-6 (50 ng/ml each) plus ruxolitinib (5 µM), as indicated. (L) Densitometric semi-quantification of p-STAT1/STAT1 (ratio). (B and D) Unpaired two-tailed Student's t-test. (E, F, J and L) One-way ANOVA with Tukey's post hoc multiple comparisons test. *P<0.05, ***P<0.001, ****P<0.0001. CTRL, control; IBA-1, ionised calcium binding adaptor molecule 1; ns, not significant; p-, phosphorylated; PE-A, phycoerythrin-area.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Immunohistochemistry, Staining, Expressing, Scratch Wound Assay Assay, Western Blot, Two Tailed Test, Control, Binding Assay

Journal: Molecular Medicine Reports

Article Title: Elevated IgG levels induce an M2-to-M1 phenotypic shift in mucosal macrophages and restrict the growth of invasive sphenoid sinus pituitary adenomas

doi: 10.3892/mmr.2026.13878

Figure Lengend Snippet: Summary graphic illustration. This illustration summarizes the proposed model during pituitary adenoma invasion. The tumor invasive front abuts an intact sphenoid sinus mucosa, forming a distinct boundary. The mucosal compartment is enriched for ionised calcium binding adaptor molecule 1-positive macrophages with an M1-like predominance and IgG-high B cells. B cell-derived IgG promotes M2-to-M1 macrophage reprogramming, while coordinated IFN-γ and IL-6 production establishes a tumor-suppressive cytokine gradient that decreases from mucosa toward the tumor core, constraining proliferation and migration via JAK-STAT1 activation. Therapeutically, anti-CD47 monoclonal antibody blocks the CD47-SIRPα ‘don't-eat-me’ axis and augments antibody-dependent cellular phagocytosis, highlighting a strategy for immune checkpoint-targeted therapy that may complement surgical management. FcR, Fc receptor; JAK, Janus kinase; mAb, monoclonal antibody; p-, phosphorylated; SIRPα, signal regulatory protein-α.

Article Snippet: RAW264.7 macrophages in the M0, M1 or M2 state were treated with IgG (10 μg/ml; cat. no. 14-4714-85; Invitrogen; Thermo Fisher Scientific, Inc.) or anti-CD47 monoclonal antibody (mAb) (10 μg/ml; cat. no. 16-0479-85; Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C for 12 h. For polarization, M1 macrophages were induced with lipopolysaccharide (100 ng/ml; cat. no. HY-D1056; MedChemExpress) plus IFN-γ (20 ng/ml; cat. no. RP01070; ABclonal Biotech Co., Ltd.) for 24 h at 37°C, whereas M2 macrophages were induced with IL-4 (20 ng/ml; cat. no. RP01161; ABclonal Biotech Co., Ltd.) for 24 h at 37°C.

Techniques: Binding Assay, Derivative Assay, Migration, Activation Assay

Journal: Open Medicine

Article Title: NTN1 regulates autophagy through the MAP1B/DAPK1 axis to ameliorate acute kidney injury in vitro

doi: 10.1515/med-2025-1374

Figure Lengend Snippet: The effects of NTN1 on the apoptosis, inflammatory levels and ROS generation in LPS-treated HK-2 cells (A–B) after the transfection of NTN1 overexpression plasmid or si-NTN1, the expression of NTN1 in the blank, OE-NC, OE-NTN1, si-NC, and si-NTN1 groups was determined by qRT-PCR. GAPDH served as the internal control. (C–D) the apoptosis rate of HK-2 cells or RPTEC/TERT1 in the OE-NC, OE-NTN1, si-NC, and si-NTN1 groups was assessed by flow cytometry. (E–G) the levels of TNF-α, IL-6 and IL-1β in the OE-NC, OE-NTN1, si-NC, and si-NTN1 groups were determined by ELISA. (H) The level of ROS generation in each group was detected by DCFH-DA reagent. The data are presented as the mean±standard deviation of three independent experiments; *** p<0.001 vs. OE-NC; ### p<0.001 vs. si-NC. Abbreviation: NTN1, netrin 1; ROS, reactive oxygen species; LPS, lipopolysaccharides; OE-NTN1, NTN1 overexpression; si-NTN1, silenced NTN1; NC, negative control; qRT-PCR, quantitative real-time PCR; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; TNF-α, tumor necrosis factor-α; IL-6, interleukin-6; ELISA, enzyme-linked immunosorbent assay; DCFH-DA, 2,7-Dichlorodi-hydrofluorescein diacetate.

Article Snippet: To assess the role of NTN1 in renal injury in vitro , HK-2 or RPTEC/TERT1 cells were transfected with NTN1 overexpression plasmid or si-NTN1 and then stimulated with 10 μg/mL lipopolysaccharide (LPS, HY-D1056, MedChemExpress, China) to construct cell injury model [ ].

Techniques: Transfection, Over Expression, Plasmid Preparation, Expressing, Quantitative RT-PCR, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Standard Deviation, Negative Control, Real-time Polymerase Chain Reaction

Journal: Open Medicine

Article Title: NTN1 regulates autophagy through the MAP1B/DAPK1 axis to ameliorate acute kidney injury in vitro

doi: 10.1515/med-2025-1374

Figure Lengend Snippet: The interaction between MAP1B and NTN1 (A) the interaction between MAP1B and NTN1 was analyzed by string website ( https://string-db.org/ ). (B–C) the ratio of p-MAP1B/MAP1B in the OE-NC, OE-NTN1, si-NC, and si-NTN1 groups was determined by western blot. GAPDH served as the internal control. The data are presented as the mean±standard deviation of three independent experiments; ** p<0.01, *** p<0.001 vs. OE-NC; # p<0.05, ## p<0.01 vs. si-NC. Abbreviation: MAP1B, microtubule associated protein 1B; NTN1, netrin 1; OE-NTN1, NTN1 overexpression; si-NTN1, silenced NTN1; NC, negative control; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: To assess the role of NTN1 in renal injury in vitro , HK-2 or RPTEC/TERT1 cells were transfected with NTN1 overexpression plasmid or si-NTN1 and then stimulated with 10 μg/mL lipopolysaccharide (LPS, HY-D1056, MedChemExpress, China) to construct cell injury model [ ].

Techniques: Western Blot, Control, Standard Deviation, Over Expression, Negative Control