Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Summary of the immunocytochemical analysis of CNHCs with antibodies against CD45, CD31, and CAIX

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques:

Journal: Journal of Translational Medicine

Article Title: Are morphological criteria sufficient for the identification of circulating tumor cells in renal cancer?

doi: 10.1186/1479-5876-11-214

Figure Lengend Snippet: Immunocytochemical analysis of CNHCs with antibodies against the RCC marker CAIX. Clusters of CNHCs cytomorphologically classified as uncertain malignant (−UMF) with cytoplasmic positive staining with antibodies against the RCC marker CAIX (A) . Clusters of CNHC-UMF and -BF without reactivity for CAIX antibodies ( B and C , respectively). A single CNHC-MF with positive cytoplasmic (D) and without staining for CAIX (E) . Single CAIX-negative CNHC-UMF and -BF ( F and G , respectively).

Article Snippet: The filters were incubated with primary antibodies directed against CAIX (rabbit IgG, NB100-417, Novus Biologicals, Littleton, USA; 3.5 μg/ml ), CD31 (mouse IgG1, M0823, Dako, Glostrup, Denmark; 1 μg/ml) or CD45 (mouse IgG1, M0855, Dako, Glostrup, Denmark; 2.4 μg/ml) diluted in Antibody Diluent (Dako, Glostrup, Denmark) for 30 min at RT followed by application of primary antibody enhancer (Dako, Glostrup, Denmark) for 15 min.

Techniques: Marker, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: An immunostimulant nanomedicine enhances radioimmunotherapy by remodeling the tumor immunosuppressive landscape after radiotherapy

doi: 10.1016/j.apsb.2025.11.012

Figure Lengend Snippet: Immunosuppression after radiotherapy promotes breast cancer progression. Tumor growth curves (A) and weights (B) of mice before and after RT ( n = 6). (C) Survival curve of mice after different treatments ( n = 6). Tumor growth curves (D) and weights (E) of mice treated with RT and RT plus α -PD-L1 ( n = 6). (F) Survival curve of mice after different treatments ( n = 6). (G) Cluster analysis of differential expression genes between untreated and RT-treated tumors 48 h post-RT ( n = 3). (H) Gene Ontology (GO) analysis of tumor tissues before and after RT (select the top 10 for each item). (I) Heat map of differentially expressed genes related to apoptosis and immune suppression ( n = 3). (J) Immunofluorescence staining images of tumor tissue in saline and RT groups (scale bar = 20 μm). (K) Quantitative analysis of tumor-infiltrating CD45 + cells ( n = 6). Representative flow cytometry images (M) and quantitative analysis (L) of tumor-infiltrating MDSCs ( n = 6). (N) The TUNEL staining of tumor section (scale: 25 μm). (O) Adenosine content detection in tumor tissue ( n = 6). Data are presented as mean ± SD. ∗∗∗∗ P < 0.0001 determined by Student’s t-test.

Article Snippet: InVivo MAb anti-mouse PD-L1 was purchased from Bioxcell (USA).

Techniques: Quantitative Proteomics, Immunofluorescence, Staining, Saline, Flow Cytometry, TUNEL Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: An immunostimulant nanomedicine enhances radioimmunotherapy by remodeling the tumor immunosuppressive landscape after radiotherapy

doi: 10.1016/j.apsb.2025.11.012

Figure Lengend Snippet: FD@ATRA-enhanced radiotherapy combined with α -PD-L1 to enhance the efficacy of large-volume tumors. (A) Schematic diagram of the therapeutic process for evaluating the anti-tumor effects in a bilateral 4T1 tumor-bearing mouse model of large volume. (B) Distant tumor growth curves of mice after different treatments ( n = 5). (C) In vitro images of the distant tumors ( n = 5). (D) In vitro distant tumors mass ( n = 5). (E) Immunohistochemical staining of CD3 in tumor tissue slices after different treatments (scale bars = 100 μm). The images below were the corresponding enlarged parts (scale bars = 25 μm). (F) Immunohistochemical staining of CD8 in tumor tissue slices after various treatments (scale bars = 100 μm). The images below were the corresponding enlarged parts (scale bars = 25 μm). Data are presented as mean ± SD. ∗∗∗ P < 0.001, and ∗∗∗∗ P < 0.0001 determined by Student’s t -test.

Article Snippet: InVivo MAb anti-mouse PD-L1 was purchased from Bioxcell (USA).

Techniques: In Vitro, Immunohistochemical staining, Staining

Journal: Theranostics

Article Title: Hypoxia-induced TGFBI maintains glioma stem cells by stabilizing EphA2

doi: 10.7150/thno.95141

Figure Lengend Snippet: TGFBI is associated with the hypoxic microenvironment in human gliomas. (A) FISH staining of TGFBI in human GBM specimens (Ivy gap datasets). Scale bars: 2 mm; enlarged image: 100 μm. (B) Volcano map showing genes highly expressed in hypoxia region (pseudopalisade and microvascular proliferation regions). Each dot represents a gene. (C) Correlation between TGFBI and mRNA expression of hypoxia-related gene in the CCGA-GBM and TGGA-GBM datasets. CC, correlation coefficient; Dot size and color represent the correlation coefficient. (D) IF staining of TGFBI (green) and two hypoxia-associated markers, HIF1α (above, red) and CA9 (bottom, red), in human GBM specimens. Scale bars: 50 μm; enlarged image: 10 μm. (E) IHC staining demonstrating the association between TGFBI and HIF1α proteins in human gliomas. AOD, Average of density; n = 58.

Article Snippet: Primary antibodies: TGFBI (Abclonal, Cat#A11222, for IB, 1:1000; for IP, 5ug; Proteintech, Cat#10188-AP, for IHC, 1:100; for IF, 1:100), CD133 (Affinity, Cat#BF0403, for IF, 1:100), SOX2 (Proteintech, Cat#66411-1-Ig, for IB, 1:1000; Santa Cruz, Cat#365823, for IF, 1:50; for IHC, 1:50), HIF1α (Proteintech, Cat#20960-1-AP, for IB, 1:1000; for IF, 1:50; for IHC 1:50), carbonic anhydrase 9 (CA9, NOVUS, Cat#NB100-417, for IF, 1:50), OLIG2 (Proteintech, Cat#66513-1-Ig, for IB, 1:1000; for IF, 1:200), Ki67 (Proteintech, Cat#27309-1-AP, for IF, 1:200), EphA2 (Abclonal, Cat#A7183, for IB,1:1000; Santa Cruz, Cat#398832, for IF, 1:50; for IHC, 1:50; for IP, 5ug), Akt (Proteintech, Cat#60203-1-Ig, for IB, 1:5000), phospho-Akt (Ser473) (Proteintech, Cat#66444-1-Ig, for IB, 1:2000; Cell Signaling Technology, Cat#4060, for IF, 1:100; for IHC, 1:50), c-MYC (Proteintech, Cat#67447-1-Ig, for IB, 1:5000; for IF,1:100; for IHC, 1:100), phospho-EphA2 (Y588) (Proteintech, Cat#30263-1-AP, for IB, 1:1000), Integrin αV (Proteintech, Cat#27096-1-AP, for IB, 1:1000), Integrin β5 (CST, Cat#3629, for IB, 1:1000), Notch1 (Cell Signaling Technology, Cat#3608, for IB, 1:1000).

Techniques: Staining, Expressing, Immunohistochemistry

Journal: Frontiers in Cell and Developmental Biology

Article Title: Garciniagifolone A, derived from Garcinia multiflora fruits, inhibits triple-negative breast cancer cells and organoids growth by targeting CA9

doi: 10.3389/fcell.2026.1767397

Figure Lengend Snippet: GA inhibits TNBC growth via targeting CA9. (a) Venn diagram showing the number of differentially expressed genes in the comparison between MDA-MB-231 and BT549. (b,c) TCGA and GEO databases show that CA9 is highly expressed in TNBC. (d) Correlation analysis between TNBC patient prognosis and CA9 expression. (e) IHC staining of TNBC patient-derived cancer tissue and adjacent tissue. Scale bar: 100 μm. (f) The quantitative analysis of CA9. (g) The protein levels of CA9 in cell lines were measured by Western blotting. (h) The predicted binding mode of GA with CA9. (i) GA binds to CA9 protein as shown by SPR. (j,k) CETSA analysis of CA9 combined with GA. (l,m) CA9 mRNA levels after GA 24 h treatment by qRT-PCR. (n,o) CA9 protein levels after GA 24 h treatment by Western blotting. (p) The related protein levels of siRNA-CA9 TNBC cells after GA 24 h treatment by Western blotting. Data are presented as mean ± SD. n = 3. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 when compared with control group.

Article Snippet: The CA9 protein (Cusabio, CSB-EP614990HUc0) was coupled to the surface of CM5 chip biosensor (Cytiva, 29104988) in 10 mM sodium acetate (pH 4.5).

Techniques: Comparison, Expressing, Immunohistochemistry, Derivative Assay, Western Blot, Binding Assay, Quantitative RT-PCR, Control

Journal: Frontiers in Cell and Developmental Biology

Article Title: Garciniagifolone A, derived from Garcinia multiflora fruits, inhibits triple-negative breast cancer cells and organoids growth by targeting CA9

doi: 10.3389/fcell.2026.1767397

Figure Lengend Snippet: Model of the anti-tumor mechanism of GA in TNBC cells. GA could bind CA9 to induce early stage autophagy by blocking PI3K/AKT/mTOR signaling pathway. Moreover, GA suppressed the autophagic flux and increased the p62 to promote ROS production, which activated NLRP3/caspase-1/GSDMD/caspase-3 related pyroptosis and apoptosis to induce TNBC cells death.

Article Snippet: The CA9 protein (Cusabio, CSB-EP614990HUc0) was coupled to the surface of CM5 chip biosensor (Cytiva, 29104988) in 10 mM sodium acetate (pH 4.5).

Techniques: Blocking Assay