Journal: Oncology Letters

Article Title: FOXP3 and CEACAM6 expression and T cell infiltration in the occurrence and development of colon cancer

doi: 10.3892/ol.2016.4439

Figure Lengend Snippet: Sequence of the primer pairs used for reverse transcription-quantitative polymerase chain reaction.

Article Snippet: The tissue slides were blocked for 10 min at room temperature (25°C), washed with phosphate-buffered saline (PBS), then incubated with the following anti-human antibodies at room temperature (25°C) for 2 h in moist dark chambers: Monoclonal mouse IgG1 against CD3 (#sc-20047), polyclonal rabbit IgG against CD4 (#sc-7219) and CD8 (#sc-7188; all from Santa Cruz Biotechnology, Inc., Dallas, TX, USA; dilution, 1:100); monoclonal mouse IgG2a against CD45RO (#ab86080; dilution, 1:10), monoclonal rabbit IgG against CEACAM6 (#ab134074; dilution, 1:400) and monoclonal mouse IgG3 against FOXP3 (#ab450; 1:50 dilution; all from Abcam, Cambridge, MA, USA).

Techniques: Sequencing

Journal: Oncology Letters

Article Title: FOXP3 and CEACAM6 expression and T cell infiltration in the occurrence and development of colon cancer

doi: 10.3892/ol.2016.4439

Figure Lengend Snippet: Representative images of CD3, CD4, CD8, CD45RO, CEACAM6 and FOXP3 staining by immunohistochemistry in normal colonic tissue, colonic adenoma and colon cancer. Images show 3,3′-diaminobenzidine tetrahydrochloride hydrate staining of positive cells. CD3, CD4, CD8 and CD45RO were immunolocalized to the membrane, CEACAM6 in the cytoplasmic and FOXP3 in the nucleus. All images were taken at ×400 scanning magnification. CD, cluster of differentiation; CEACAM6, carcinoembryonic antigen-related cell adhesion molecule 6; FOXP3, forkhead box P3.

Article Snippet: The tissue slides were blocked for 10 min at room temperature (25°C), washed with phosphate-buffered saline (PBS), then incubated with the following anti-human antibodies at room temperature (25°C) for 2 h in moist dark chambers: Monoclonal mouse IgG1 against CD3 (#sc-20047), polyclonal rabbit IgG against CD4 (#sc-7219) and CD8 (#sc-7188; all from Santa Cruz Biotechnology, Inc., Dallas, TX, USA; dilution, 1:100); monoclonal mouse IgG2a against CD45RO (#ab86080; dilution, 1:10), monoclonal rabbit IgG against CEACAM6 (#ab134074; dilution, 1:400) and monoclonal mouse IgG3 against FOXP3 (#ab450; 1:50 dilution; all from Abcam, Cambridge, MA, USA).

Techniques: Staining, Immunohistochemistry, Membrane

Journal: Oncology Letters

Article Title: FOXP3 and CEACAM6 expression and T cell infiltration in the occurrence and development of colon cancer

doi: 10.3892/ol.2016.4439

Figure Lengend Snippet: Immunohistochemical staining scores of T cell subsets in normal colonic tissue, colonic adenoma, stage I–II and stage III–IV cancer tissue. (A) CD3, (B) CD4, (C) CD8, and (D) CD45RO staining scores (n=12 for the normal group, n=38 for the adenoma group, n=40 for the stage I–II group, n=38 for the stage III–IV group). Whiskers represent the range and boxes represent the quartile values. *P<0.05 vs. normal group; ^ P<0.05 vs. adenoma group; # P<0.05 vs. stage I–II cancer group. CD, cluster of differentiation.

Article Snippet: The tissue slides were blocked for 10 min at room temperature (25°C), washed with phosphate-buffered saline (PBS), then incubated with the following anti-human antibodies at room temperature (25°C) for 2 h in moist dark chambers: Monoclonal mouse IgG1 against CD3 (#sc-20047), polyclonal rabbit IgG against CD4 (#sc-7219) and CD8 (#sc-7188; all from Santa Cruz Biotechnology, Inc., Dallas, TX, USA; dilution, 1:100); monoclonal mouse IgG2a against CD45RO (#ab86080; dilution, 1:10), monoclonal rabbit IgG against CEACAM6 (#ab134074; dilution, 1:400) and monoclonal mouse IgG3 against FOXP3 (#ab450; 1:50 dilution; all from Abcam, Cambridge, MA, USA).

Techniques: Immunohistochemical staining, Staining

Journal: Oncology Letters

Article Title: FOXP3 and CEACAM6 expression and T cell infiltration in the occurrence and development of colon cancer

doi: 10.3892/ol.2016.4439

Figure Lengend Snippet: Expression of CD3, CD4, CD8 and CD45RO mRNA in colon tissues of each group. (A) CD3, (B) CD4, (C) CD8 and (D) CD45RO mRNA levels (n=6 for the normal group, n=10 for the adenoma group, n=10 for the stage I–II group, n=9 for the stage III–IV group). *P<0.05 vs. normal group; ^ P<0.05 vs. adenoma group; # P<0.05 vs. stage I–II cancer group. CD, cluster of differentiation.

Article Snippet: The tissue slides were blocked for 10 min at room temperature (25°C), washed with phosphate-buffered saline (PBS), then incubated with the following anti-human antibodies at room temperature (25°C) for 2 h in moist dark chambers: Monoclonal mouse IgG1 against CD3 (#sc-20047), polyclonal rabbit IgG against CD4 (#sc-7219) and CD8 (#sc-7188; all from Santa Cruz Biotechnology, Inc., Dallas, TX, USA; dilution, 1:100); monoclonal mouse IgG2a against CD45RO (#ab86080; dilution, 1:10), monoclonal rabbit IgG against CEACAM6 (#ab134074; dilution, 1:400) and monoclonal mouse IgG3 against FOXP3 (#ab450; 1:50 dilution; all from Abcam, Cambridge, MA, USA).

Techniques: Expressing

Journal: Indian Journal of Dermatology

Article Title: ERK/MEK Pathway Regulates Th17 Cell Differentiation in Patients with Pemphigus Vulgaris

doi: 10.4103/ijd.ijd_924_22

Figure Lengend Snippet: Abnormality of Th17 cells in the peripheral blood of PV patients. (a) The proportion of Th17 cells (CD3+, CD8-, IL-17A+) to CD4+ T cells (CD3+, CD8-) between healthy controls and the PV patients, which was detected by flow cytometry and illustrated in a histogram. (b) The serum levels of IL-17 were detected using ELISA. (c) Relative mRNA levels of RORγt in CD4+ T cells were measured by qPCR. (d) Immunoblot analysis of Th17 lineage-associated proteins (STAT3, p-STAT3, RORγt and IL-17) in CD4+ T cells. The relative intensity was normalised to the levels of GAPDH and showed in a histogram. (e) Immunofluorescence staining of accumulated p-STAT3 in Th17 cells. p-STAT3 was stained in red, and nuclei were stained by DAPI in blue. Magnification: 400×. Scale bar: 50 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: CS1001 and CS1002; Multisciences (Lianke) Biotech, Hangzhou, China) at 37°C in a humidified incubator with 5% CO2 for 4 h. After being stained with PE-labelled anti-human CD3 antibody and FITC-labelled anti-human CD8a antibody (Catalogue Nos.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Immunofluorescence, Staining

Journal: Indian Journal of Dermatology

Article Title: ERK/MEK Pathway Regulates Th17 Cell Differentiation in Patients with Pemphigus Vulgaris

doi: 10.4103/ijd.ijd_924_22

Figure Lengend Snippet: Inhibition of the ERK signalling prevented Th17 cell differentiation in PV. The inhibition was performed in vitro by the addition of PD98059. DMSO was used as a reagent control. (a) The proportion of Th17 cells (CD3+, CD8-, IL-17A+) to CD4+ T cells (CD3+, CD8-), which was detected by flow cytometry. (b) The mRNA levels of RORγt were determined using qPCR. (c) The IL-17 levels in cell culture supernatant were detected by ELISA. *, P < 0.05; **, P < 0.01

Article Snippet: CS1001 and CS1002; Multisciences (Lianke) Biotech, Hangzhou, China) at 37°C in a humidified incubator with 5% CO2 for 4 h. After being stained with PE-labelled anti-human CD3 antibody and FITC-labelled anti-human CD8a antibody (Catalogue Nos.

Techniques: Inhibition, Cell Differentiation, In Vitro, Control, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Journal for Immunotherapy of Cancer

Article Title: Loss of ADAR1 in macrophages in combination with interferon gamma suppresses tumor growth by remodeling the tumor microenvironment

doi: 10.1136/jitc-2023-007402

Figure Lengend Snippet: ADAR1 loss in macrophage with IFN-γ treatment affects the secretion of key cytokines through PKR/EIF2α signaling. (A, C) Human XL cytokine arrays for detecting differential factors between THP-1 cells with scrambled shRNA and shADAR1#1 under the treatment of IFN-γ (A) and between THP-1 cells with empty vector and WT ADAR , co-cultured with A549 (C). (B, D) Bar plots showing the expression levels of differential factors on ADAR1 knockdown (B) and ADAR1 overexpression (D). (E) Venn gram showing key cytokines identified by ADAR1 knockdown and overexpression experiments and their potential effects on the tumor microenvironment. (F) RT-qPCR-based mRNA expression levels of ADAR , CCL20 , GDF15 , IFN-G , IL-18 , IL-18BP , and HAVCR2 in different THP-1 cells (transfected with scrambled shRNA, shADAR1#1 or shADAR1#2) with IFN-γ treatment. β-actin was used as an internal control. (G) RT-qPCR-based mRNA expression levels of Ccl20, Gdf15, Il-18, Il-18bp , and Havcr2 in BMDMs from C57BL/6 mice ( Adar fl/fl and Adar fl/fl Lyz2 Cre ) treated with IFN-γ. Gapdh was used as an internal control. (H) Immunofluorescent staining for anti-dsRNA (J2) in THP-1 cells treated with IFN-γ. RNase III treatment was used as the negative control for the dsRNA signal. Scale bars, 10 µm. (I) Western blot showing the protein expression of p-PKR Thr446/Thr451 , PKR, p-EIF2α Ser51 , and EIF2α protein expression in THP-1 cells transfected with scrambled shRNAs and treated with poly I:C (1 µg/mL). (J) Western blot showing the protein expression of p-PKR Thr446/Thr451 , PKR, p-EIF2α Ser51 , and EIF2α expression in THP-1 cells (transfected with scrambled, shADAR1#1, or shADAR1#2) with IFN-γ treatment. (K) Western blot showing the protein expression levels of p-PKR Thr446/Thr451 , PKR, p-EIF2α Ser51 , and EIF2α protein expression in THP-1(shADAR1#1) cells with IFN-γ after pretreatment with or without 2-AP. (L) ELISA quantitative measurement of Ccl20, Gdf15, Il-18, Il-18bp, and Tim-3 from conditioned media of different BMDMs ( Adar fl/fl , Adar fl/fl Lyz2 Cre , and Adar fl/fl Lyz2 Cre with 2-AP) treated with IFN-γ. (M) Dual luciferase reporter assays for the 5'UTR activities of CCL20 , HAVCR2, IL-18 , and IL-18- Mutant. Data are normalized to renilla luciferase. (B, D, F, G, L, and M) Data are shown as mean±SD, and p values are based on unpaired Student’s t-test. (I, J and K) β-actin was used as the loading control. ADAR, adenosine deaminases acting on RNA; BMDMs, bone marrow-derived macrophages; dsRNAs, double-stranded RNAs; IFN, interferon; IL, interleukin; mRNA, messenger RNA; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; WT, wild type; 2-AP, 2-aminopurine.

Article Snippet: Anti-ADAR1 antibody (Santa Cruz, sc-73408), anti-ADAR1 antibody (Santa Cruz, sc-271854), anti-V5 antibody (Invitrogen, R960-251), anti-β-actin antibody (EnoGene, E12-041-1), anti-CD31 antibody (Abcam, ab28364), anti-PKR (phospho T446) antibody (Abcam, ab32036), anti-PKR (phospho T451) antibody (Abcam, ab81303), anti-PKR antibody (ProteinTech, 18 244–1-AP), anti-CD8a antibody (Invitrogen, 14-0081-85), anti-EIF2α antibody (ProteinTech, 11 170–1-AP), anti-EIF2α (phospho Ser51) antibody (Abmart, TA3087S), anti-J2 antibody (SCICONS, 10010200), anti-CD8a antibody (Cell Signaling Technology, 98941), anti-CD28 antibody (Abmart, TA0014S), anti-CD3 antibody (Invitrogen, 14-0032-82), PE anti-mouse CD8a antibody (Elabscience, E-AB-F1104D), APC anti-mouse Perforin Antibody (BioLegend, S16009B), APC Rat IgG2a, κ Isotype Control (Elabscience, E-AB-F09832E), APC anti-human/mouse Granzyme B Recombinant Antibody (BioLegend, 372204), APC mouse IgG1, κ Isotype Control (Elabscience, E-AB-F09792E), APC anti-mouse/human CD11b antibody (BioLegend, 101212), FITC anti-mouse F4/80 antibody (BioLegend, 123108), Alexa Fluor 488 AffiniPure Donkey Anti-Rat IgG (H+L) (Jackson, 712-545-150), Alexa Fluor 594 AffiniPure Donkey Anti-Rabbit IgG (H+L) (Jackson, 711-585-152).

Techniques: shRNA, Plasmid Preparation, Cell Culture, Expressing, Knockdown, Over Expression, Quantitative RT-PCR, Transfection, Control, Staining, Negative Control, Western Blot, Enzyme-linked Immunosorbent Assay, Luciferase, Mutagenesis, Derivative Assay, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Materials Today Bio

Article Title: A pH/MMP-9 smart dual-responsive liposome GBE@LP co-delivers and controls the release of GB1107/BMS1166/Enzalutamide for liver cancer immunotherapy

doi: 10.1016/j.mtbio.2025.101801

Figure Lengend Snippet: Graphical illustration of the immunization strategy of GBE@LP for liver cancer treatment. GBE@LP has prolonged circulation in the bloodstream due to surface PEG and selectively accumulates in liver cancer tissues through EPR effect. Meanwhile, DOPE and TGMS give it the property to be released in response to low pH and MMP-9 overexpression in liver cancer microenvironment. GBE@LP delivers drugs into liver cancer, converting uninfiltrated "cold" tumors into highly infiltrated "hot" tumors, while reversing CD8 + T-cell depletion and enhancing cytotoxicity. These effects ultimately enhance immunotherapy for liver cancer.

Article Snippet: After deparaffinization and rehydration, the sections were microwaved in 0.01M sodium citrate (pH 6) for 8 min, followed by blocking with 5 % bovine serum albumin (BSA) for 1 h. The sections were then incubated with anti-mouse CD8, anti-mouse PD-1, and anti-mouse AR antibodies (all from Proteintech) at 4 °C for 12 h. After washing, the sections were incubated with Dylight 649 (Goat Anti-Mouse IgG) and Dylight 488 (Goat Anti-Rabbit IgG) antibodies (all from Abbkine) for 1 h, stained with DAPI for 5 min, and then mounted.

Techniques: Over Expression

Journal: Materials Today Bio

Article Title: A pH/MMP-9 smart dual-responsive liposome GBE@LP co-delivers and controls the release of GB1107/BMS1166/Enzalutamide for liver cancer immunotherapy

doi: 10.1016/j.mtbio.2025.101801

Figure Lengend Snippet: Liver cancer is a "cold" tumor with high Gal-3 expression. (A) Immunofluorescence images of CD8 + T cells (red) and DAPI (blue) in liver cancer tissues of tumor-bearing mice. (B) Immunohistochemical images of cd8 + T cells in patients with hepatocellular carcinoma (HCC), renal cell carcinoma (RCC), and lung cancer (LC) from the HPA database. (C) Expression of Gal-3 in different human cancers was analyzed by HPA database. (D) The expression of Gal-3 in mouse liver cancer tissues and paracancerous tissues was detected by western blotting. (E) The expression of CD8 in the control group and GB1107 treatment group was detected by western blotting. (F) Immunofluorescence images of CD8 + T cells (green), PD-1 (red), and DAPI (blue) in liver cancer tissues and paracancerous tissues of tumor-bearing mice. (G) Immunofluorescence images of CD8 + T cells (green), AR (red), and DAPI (blue) in liver cancer tissues and paracancerous tissues of tumor-bearing mice.

Article Snippet: After deparaffinization and rehydration, the sections were microwaved in 0.01M sodium citrate (pH 6) for 8 min, followed by blocking with 5 % bovine serum albumin (BSA) for 1 h. The sections were then incubated with anti-mouse CD8, anti-mouse PD-1, and anti-mouse AR antibodies (all from Proteintech) at 4 °C for 12 h. After washing, the sections were incubated with Dylight 649 (Goat Anti-Mouse IgG) and Dylight 488 (Goat Anti-Rabbit IgG) antibodies (all from Abbkine) for 1 h, stained with DAPI for 5 min, and then mounted.

Techniques: Expressing, Immunofluorescence, Immunohistochemical staining, Western Blot, Control

Journal: Materials Today Bio

Article Title: A pH/MMP-9 smart dual-responsive liposome GBE@LP co-delivers and controls the release of GB1107/BMS1166/Enzalutamide for liver cancer immunotherapy

doi: 10.1016/j.mtbio.2025.101801

Figure Lengend Snippet: GBE@LP remodeling the immunosuppressive microenvironment of liver cancer. (A) Flow cytometry scatter graph and (C) Flow cytometry statistical plot of CD8 + T cells infiltration in the tumors of different treatment groups. (B) Flow cytometry histogram and (D) Flow cytometry statistical plot of CD3 + T cells infiltration in the spleen of different treatment groups. (n = 3, p ∗ < 0.05, p ∗∗ < 0.01, p ∗∗∗ < 0.001, p ∗∗∗∗ < 0.0001).

Article Snippet: After deparaffinization and rehydration, the sections were microwaved in 0.01M sodium citrate (pH 6) for 8 min, followed by blocking with 5 % bovine serum albumin (BSA) for 1 h. The sections were then incubated with anti-mouse CD8, anti-mouse PD-1, and anti-mouse AR antibodies (all from Proteintech) at 4 °C for 12 h. After washing, the sections were incubated with Dylight 649 (Goat Anti-Mouse IgG) and Dylight 488 (Goat Anti-Rabbit IgG) antibodies (all from Abbkine) for 1 h, stained with DAPI for 5 min, and then mounted.

Techniques: Flow Cytometry

Journal: Materials Today Bio

Article Title: A pH/MMP-9 smart dual-responsive liposome GBE@LP co-delivers and controls the release of GB1107/BMS1166/Enzalutamide for liver cancer immunotherapy

doi: 10.1016/j.mtbio.2025.101801

Figure Lengend Snippet: (A) Immunofluorescence images of CD8 + T cells (green) and DAPI (blue) in different treatment groups. Scale bars, 1000 μm (top), 50 μm (bottom).

Article Snippet: After deparaffinization and rehydration, the sections were microwaved in 0.01M sodium citrate (pH 6) for 8 min, followed by blocking with 5 % bovine serum albumin (BSA) for 1 h. The sections were then incubated with anti-mouse CD8, anti-mouse PD-1, and anti-mouse AR antibodies (all from Proteintech) at 4 °C for 12 h. After washing, the sections were incubated with Dylight 649 (Goat Anti-Mouse IgG) and Dylight 488 (Goat Anti-Rabbit IgG) antibodies (all from Abbkine) for 1 h, stained with DAPI for 5 min, and then mounted.

Techniques: Immunofluorescence