Journal: Journal of Cancer Research and Clinical Oncology

Article Title: Regulatory T cells inhibit FoxP3 to increase the population of tumor initiating cells in hepatocellular carcinoma

doi: 10.1007/s00432-024-05892-2

Figure Lengend Snippet: ‘Stemness’ of HCC-LM3 cells was enhanced after co-cultured with Tregs. ( a ) Isolation and expansion of CD4 + CD25 + CD127- Tregs. ( b ) Flow cytometry analysis showing high expression of CD4, CD25, and FoxP3 in Tregs. ( c ) Co-culture of encapsulated HCC cells and Tregs. ( d ) Immunofluorescence staining of CD133 in HCC cells before and after co-culture, and ( e ) HCC TIC (CD133 positive cells) ratio before and after co-culture, student’s t-test, * P < 0.05, compared with the control group. ( f ) RT-qPCR analysis of HCC TICs-related genes and ( g ) EMT-related genes, n = 3, student’s t-test, * P < 0.05, compared with the control group. ( h ) Western blot analysis of β-catenin and c-Myc in HCC cells before and after co-culture

Article Snippet: Tregs were labeled with FITC Mouse Anti-Human CD4 (1:5) (561,005, BD Biosciences, Franklin Lakes, NJ, USA), PE Mouse Anti-Human CD25 (1:5) (555,432, BD Biosciences) and Alexa Fluor ® 647 Mouse anti-Human FoxP3 (1:20) (561,184, BD Biosciences) antibodies for 30 min on ice, followed by washing with phosphate buffered saline (PBS) (Gibco), FITC Mouse IgG1 (555,748, BD Biosciences), PE Mouse IgG1 (555,749, BD Biosciences), Alexa Fluor ® 647 Mouse IgG1 (557,732, BD Biosciences) were used as isotype controls.

Techniques: Cell Culture, Isolation, Flow Cytometry, Expressing, Co-Culture Assay, Immunofluorescence, Staining, Control, Quantitative RT-PCR, Western Blot

Journal: iScience

Article Title: Human skin CD141 + dendritic cells regulate cutaneous immunity via the neuropeptide urocortin 2

doi: 10.1016/j.isci.2023.108029

Figure Lengend Snippet: UCN2 protects from alloimmune cell-mediated human skin inflammation in vivo (A) qPCR CRHR2 expression in cell populations isolated from human skin and blood, presented as arbitrary units (AU). (B) T cell suppression assay from sorted CD25 hi T cells from co-cultures with either CD1c + DDCs or CD141 + DDCs in the presence of vehicle control, CRHR2 antagonist (aCRHR2) alone, or together with UCN2 peptide. (C‒F) Experimental strategy to assess contribution of intradermally injected vehicle (PBS), aCRHR2 or UCN2 into human skin grafted NSG mice receiving intravenous PBS or allogeneic CD4 + T cells. Representative fields from stained skin grafts for (D) epidermal keratinocyte expression of the proliferation marker Ki67 (arrows), (E) epidermal CD3 + T cell infiltration (arrows), and (F) dermal FoxP3 + CD3 + T cells (arrows). Dashed lines indicate epidermal-dermal junction. Bars: 100μm. (G) Quantitative histological analysis of at least three independent visual fields per skin graft. Lines represent the mean ± SEM. (n = 3–6 animals per treatment group). Results are combined data from two independent experiments. One-way ANOVA test (B, G), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 See also Figure S6 .

Article Snippet: Mouse monoclonal anti-human CD25-PE (Clone: 2A3) , BD , Cat #341011.

Techniques: In Vivo, Expressing, Isolation, Suppression Assay, Injection, Staining, Marker

Journal: iScience

Article Title: Human skin CD141 + dendritic cells regulate cutaneous immunity via the neuropeptide urocortin 2

doi: 10.1016/j.isci.2023.108029

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-human CD25-PE (Clone: 2A3) , BD , Cat #341011.

Techniques: Purification, Recombinant, Staining, Concentration Assay, Sterility, Multiplex Assay, Labeling, Expressing, Transfection, Software