Journal: Cancer Research Communications

Article Title: STAG2 Expression is Associated with Adverse Survival Outcomes and Regulates Cell Phenotype in Muscle-invasive Bladder Cancer

doi: 10.1158/2767-9764.CRC-22-0155

Figure Lengend Snippet: STAG2 loss augments cisplatin treatment in MIBC. A, Kaplan–Meier OS (left) and PFS (right) survival of MIBC TMA patient samples treated with cisplatin, stratified into STAG2-low (H-score 0–50, n = 12) and -high (H-score 50–300, n = 20) groups. P = 0.125 for OS, P = 0.036 for PFS; P value computed using log-rank test. B, Scanned images of crystal violet-stained HB-CLS-1, TCC-SUP, T24, and BO2 colonies after 24 hours of treatment and 14 days of growth. Left: control, shSTAG2-1, and shSTAG-2 clonal populations treated with vehicle. Right: populations treated with 0.1 μg/mL (TCC-SUP, HB-CLS-1), 0.25 μg/mL (BO2) or 0.5 μg/mL (T24) cisplatin. Images are representative of three individual experiments. C, Surviving fraction of cells relative to untreated control for HB-CLS-1, TCC-SUP, T24, and BO2 cell lines with and without cisplatin treatment. P values determined by unpaired Student t test. Comparisons made between untreated shSTAG2-1/2 and untreated controls, and shSTAG2-1/2+CDDP to control+CDDP. Data represented as mean + SEM. CDDP: cisplatin. Individual comparisons made using unpaired Student t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: We utilized four cell lines with STAG2 KD via shSTAG2-1/2 (HB-CLS-1, TCC-SUP, T24, and BO2) in addition to UM-UC-3 cells with STAG2 overexpression.

Techniques: Staining, Control

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Phenotype of Vγ2 + T cells in STx and ATx G8 TCR-γ/δ transgenic mice. Data show proportion of Vγ2 + cells expressing different levels of ( A ) CD45RB, ( B ) HSA, ( C ) CD44, and ( D ) CD62L in pooled LNs (▪) or spleen ( ) of STx mice, or pooled LN ( ) or spleen (□) of ATx mice. Data represent mean values ± SD for three (STx) or seven (ATx) mice that had been given BrdU for 4 d.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Transgenic Assay, Expressing

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Turnover of T cells in TCR-γ/δ transgenic mice. ( A ) Total number of Vγ2 + T cells in spleen plus pooled LNs of STx versus ATx G8 TCR-γ/δ transgenic mice. Mean cell numbers (± SD) were calculated based on total cell yields and percentage of Vγ2 + cells in 10 STx and 12 ATx mice. ( B ) BrdU labeling of total Vγ2 + T cells in pooled LNs in STx (▴) and ATx (▪) G8 TCR-γ/δ transgenic mice. Mice were given BrdU in their drinking water, and Vγ2 + T cells in pooled LNs were analyzed at various times. Data represent mean values ± SD for two to seven mice for each time point.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Transgenic Assay, Labeling

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Kinetics of BrdU labeling of Vγ2 + cells in STx versus ATx G8 TCR-γ/δ transgenic mice. Mice were given BrdU continuously in their drinking water and pooled LN cells were analyzed at the indicated time points. Data show BrdU labeling of subsets of Vγ2 + cells expressing low (lo), intermediate (int), or high (hi) levels of (from top to bottom) CD44, CD45RB, CD62L or HSA in STx ( left , closed symbols ) or ATx ( right , open symbols ) TCR-γ/δ transgenic mice. Data represent mean values ± SD of two to seven mice per point.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Labeling, Transgenic Assay, Expressing

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Phenotype of γ/δ recent thymic emigrants. STx ( left ) or ATx ( right ) TCR-γ/δ transgenic mice were given BrdU for 2 d. Dot plots show representative staining of Vγ2 + pooled LN cells.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Transgenic Assay, Staining

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Pulse-chase BrdU labeling of Vγ2 + T cells in STx G8 TCR-γ/δ transgenic mice. Mice were given BrdU for 14 d (▪), or given BrdU for 14 d followed by normal drinking water for 28 d ( ). The percentage of BrdU + cells in pooled LNs is shown for ( A ) total Vγ2 + T cells; ( B ) Vγ2 + T cells expressing intermediate or high levels of CD45RB; ( C ) Vγ2 + T cells expressing low or high levels of HSA; ( D ) Vγ2 + T cells expressing low, intermediate, or high levels of CD44; and ( E ) Vγ2 + T cells expressing low or high levels of CD62L. Data represent mean values ± SD for 7 (14 d BrdU) or 2 (14 d BrdU plus 28 d normal water) mice.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Pulse Chase, Labeling, Transgenic Assay, Expressing

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Turnover of Vγ2 + thymocytes in G8 TCR-γ/δ transgenic mice. STx TCR-γ/δ transgenic mice were given BrdU for 2 (▪) or 21 ( ) d and Vγ2 + thymocytes were analyzed. ( A ) Total Vγ2 + thymocytes. ( B ) Vγ2 + CD45RB thymocyte subsets. ( C ) Vγ2 + HSA thymocyte subsets. ( D ) Vγ2 + CD44 thymocyte subsets. ( E ) Vγ2 + CD62L thymocyte subsets. Data represent mean values ± SD for two to three mice per point.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Transgenic Assay

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Phenotype of γ/δ cells activated in vivo. Vγ2 + T cells were purified from G8 TCR-γ/δ transgenic mice and injected into either β 2 m − ( upper ) or B6 ( lower ) mice; recipients were treated with anti–NK-1.1 antibody and irradiated before adoptive transfer (see Materials and Methods). After injection of Vγ2 + cells, recipients were given BrdU for 4 d. Dot plots show staining of spleen cells from representative mice.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: In Vivo, Purification, Transgenic Assay, Injection, Irradiation, Adoptive Transfer Assay, Staining

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: T cell turnover and phenotype in B6 mice. ( A ) Percentage of Vγ2 + T cells expressing low, intermediate, or high levels of CD44 in STx and ATx mice. ( B ) Proportion of Vγ2 + and CD4 + cells expressing low, intermediate, or high levels of CD44 in ATx mice. ( C ) BrdU labeling of total, CD44 lo , CD44 int and CD44 hi Vγ2 + T cells in STx and ATx B6 mice given BrdU for 7 d. ( D ) Kinetics of BrdU labeling of total Vγ2 + T cells in ATx mice. ( E ) Comparison of BrdU labeling of Vγ2 + T cells versus total TCR-γ/δ + T cells in ATx mice given BrdU for 14 d. ( F ) BrdU labeling of Vγ2 + and CD4 + T cells expressing different levels of CD44 in ATx mice given BrdU for 21 d. Data shown are for pooled LN cells and represent mean values ± SD for two to three mice per point.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Expressing, Labeling

Journal: The Journal of Experimental Medicine

Article Title: Lifespan of γ/δ T Cells

doi:

Figure Lengend Snippet: Phenotype of Vγ2 + RTEs in B6 mice. STx ( left ) or ATx ( right ) B6 mice were given BrdU for 4 d ( upper ) or 7 d ( lower ). Dot plots show representative staining of Vγ2 + pooled LN cells. The percentage of cells falling within the boxed areas is indicated.

Article Snippet: Single cell suspensions were made from pooled LNs, thymus, or spleen and were surface stained using the following antibodies: Anti-Vγ2-PE ( PharMingen , San Diego, CA); anti-CD4-PE (Collaborative Biomedical Products Co., New Bedford, MA), pan anti–TCR-γ/δ–PE ( PharMingen ); anti-CD45RB-biotin ( PharMingen ); anti-HSA-biotin (J11d); anti-CD62L-biotin (Mel-14); and anti-CD44-biotin (IM7.8.1).

Techniques: Staining