Journal: Gut microbes

Article Title: CD44 is a macrophage receptor for TcdB from Clostridioides difficile that via its lysine-158 succinylation contributes to inflammation.

doi: 10.1080/19490976.2025.2506192

Figure Lengend Snippet: Figure 3. CD44 functions as an independent receptor for TcdB/FBD in macrophages. (a – c) expression of known receptors (FZD1/2/7, CSPG4, PVRL3) in WT versus CD44-KO macrophages analyzed by (a) immunoblotting and (b, c) q-PCR. (d – g) competitive binding assays: (d, e) FZD1/2/7, CSPG4, and PVRL3 did not inhibit CD44 binding of (d) TcdB or (e) FBD, while (f, g) CD44 similarly did not interfere with TcdB/FBD binding to other receptors. (ELISA protocol: wells coated with 1 μM target protein [CD44-ECD or other receptors] were incubated with 1 μM TcdB/FBD, followed by competitor proteins. Binding was quantified by absorbance measure ment). (h – k) IL-1β (h, i) and IL-6 (j, k) levels in THP-1-Mφ treated with TcdB/FBD (100 pM) and receptor proteins (1 μM) for 24 h. All data in parts (b – k) are shown as the mean ± SEM (n = 3). ((ns: p > 0.05, **p < 0.01, &p < 0.001, #p < 0.0001).

Article Snippet: The other Primary antibodies used in this study were as follows: (Cat# 22915–1-AP), CSPG4 (Cat# 55027–1-AP), PVRL3 (Cat# 11213–1-AP), SUCLG2 (Cat# 14240–1-AP), FZD2 (Cat# 24272–1-AP), and FZD7 (Cat# 16974–1-AP) from Proteintech Group (Wuhan, China); FZD1 (Cat# PA5–86484) from Thermo Fisher Scientific; Flotillin-1 (Cat# ab41927) from Abcam (Cambridge, UK).

Techniques: Expressing, Western Blot, Binding Assay, Enzyme-linked Immunosorbent Assay, Incubation

Journal: Diagnostics

Article Title: Prognostic and Predictive Significance of B7-H3 and CD155 Expression in Gastric Cancer Patients

doi: 10.3390/diagnostics15212695

Figure Lengend Snippet: Fold change expression levels of B7-H3 and CD155 across individual gastric cancer cases. A fold change greater than 1 was interpreted as positive or upregulated gene expression, where fold regulation was considered equivalent to the fold change value. A fold change greater than 2 was classified as moderate to high upregulation in gene expression.

Article Snippet: For B7-H3, a recombinant rabbit monoclonal IgG antibody (HUABIO, Woburn, MA, USA), clone: HA721245; dilution 1:5000) was applied, while for CD155, a recombinant rabbit monoclonal IgG antibody (Proteintech, Rosemont, IL, USA, clone: 241405F5; dilution 1:1500) was used.

Techniques: Expressing, Gene Expression

Journal: Diagnostics

Article Title: Prognostic and Predictive Significance of B7-H3 and CD155 Expression in Gastric Cancer Patients

doi: 10.3390/diagnostics15212695

Figure Lengend Snippet: Comparison of B7-H3 and CD155 fold-Change Expression According to Metastatic Status (M0 vs. M1). Fold change was calculated by dividing the normalized gene expression in the test sample by the normalized gene expression in the control sample. Fold regulation represents the biologically relevant direction and magnitude of change derived from the fold change value.

Article Snippet: For B7-H3, a recombinant rabbit monoclonal IgG antibody (HUABIO, Woburn, MA, USA), clone: HA721245; dilution 1:5000) was applied, while for CD155, a recombinant rabbit monoclonal IgG antibody (Proteintech, Rosemont, IL, USA, clone: 241405F5; dilution 1:1500) was used.

Techniques: Comparison, Expressing, Gene Expression, Control, Derivative Assay

Journal: PLoS ONE

Article Title: Necl-4/SynCAM-4 Is Expressed in Myelinating Oligodendrocytes but Not Required for Axonal Myelination

doi: 10.1371/journal.pone.0064264

Figure Lengend Snippet: A. Western immuneblotting of brain tissues (from P10 wild-type and Necl-4 homozygous mice) with anti-Necl-1, anti-Necl-2 and anti-Necl-3 antibodies. β-actin was the internal control. B. Statistical analysis on the relative expression level of Necl1, Necl2 and Necl3 with Student's t-test (n = 3. Necl1, p = 0.70. Necl2, p = 0.72. Necl3, p = 0.69). Error bar, standard deviation.

Article Snippet: 30 mg protein from control and mutant tissues was loaded for SDS-PAGE electrophoresis and subsequently detected with anti-Necl-1 (developed in Peking Union Medical University), anti-Necl-2 (Proteintech, Cat# 14335-1-AP), anti-Necl-3 (Abcam Inc, Cat# ab133393) and anti-Necl-4 (UC Davis/NIH NeuroMab Facility, Cat#73-247), and mouse anti-β-actin (Sigma, Cat# A5316) antibodies according to the standard protocol.

Techniques: Western Blot, Expressing, Standard Deviation

Journal: Journal for Immunotherapy of Cancer

Article Title: Interactions between LAMP3+ dendritic cells and T-cell subpopulations promote immune evasion in papillary thyroid carcinoma

doi: 10.1136/jitc-2024-008983

Figure Lengend Snippet: The recruitment of LAMP3 + DCs by tumor cells promotes PTC clinical progression. Heatmaps (A) and circle (B) plots show the comparison of interaction quantity and interaction strength between thyrocytes and cDCs between non-progressive PTC and progressive PTC. Red indicated that the increase of communication in the latter. (C) The interaction between thyrocytes and cDCs is more numerous and stronger in Group B. (D) Summary of selected ligand-receptor interactions between thyrocytes and cDC-C3 cells in the two groups. (E) Circle plots showing the interaction between NECTIN3-NECTIN2 ligand-receptor pairs in the cDCs and thyrocytes. (F) The expression level of S100A2 is positively correlated with LAMP3. (G) Representative immunofluorescence images illustrating the interaction between thyrocytes and cDC_C3 in two groups (A1 and B9). The small panels show the magnification of the selected region highlighted in red. Scale bars correspond to 50 µm in the large panel. (H) Schematic showing the crosstalk among thyrocytes, LAMP3 + DCs, CD8 + T cells, and Tregs involved in the recruitment of immune cells and the formation of an immunosuppressive microenvironment in the progressive PTC. cDC. conventional DC; DC, dendritic cell; CTLA-4, cytotoxic T-lymphocyte associated protein 4; ICAM, intercellular adhesion molecule 1; LAMP3, lysosomal associated membrane protein 3; PTC, papillary thyroid cancer; SPN, sialophorin; TME, tumor microenvironment; Treg, regulatory T cell.

Article Snippet: The following antibodies were used in the current study: primary antibodies against DC_LAMP/CD208 (1:400, CST, Cat#47778), CD8A (1:100 dilution; Abcam, Cat#ab217344), TIGIT (1:1,000, Cell Signaling Technology, Cat#99567T), NECTIN2 (1:400, Cell Signaling Technology, Cat#95333T), NECTIN3 (1:500, R&D, Cat#AF3064-SP), CCL17 (1:1,000, Abcam, Cat#ab195044), KRT19 (1:2,000, Abcam, Cat#ab76539), FOXP3 (1:2,000, Abcam, Cat#ab215206), or CCR4 (1:2,000, Novus Biologicals, Cat#NBP1-86584).

Techniques: Comparison, Expressing, Immunofluorescence, Membrane