pdl2  (Miltenyi Biotec)

Journal: iScience

Article Title: PARP1-NFATc1-PD1 pathway of maturation and stability of CD8 + T cells is beneficial against chronic Trypanosoma cruzi infection

doi: 10.1016/j.isci.2025.112926

Figure Lengend Snippet: Antigen presenting cells (APC) profile in mice infected with T. cruzi (±PARP1) WT and Parp1 −/− mice were euthanized at 150-days post-infection (controls: non-infected) and splenocytes were analyzed by flow cytometry. (A–F) CD45 + leukocytes were examined for surface expression of CD11c, CD11b, Ly6G, and Ly6C antigens to distinguish APC subsets. The tSNE plots visualizing color-coded APC subsets (A) and percent frequencies of myeloid cells (B), monocytes (C), Mφ (D), neutrophils (E), and dendritic cells (F) in all groups of mice are shown. (G–I) Splenic leukocytes were further analyzed for surface expression of activation markers. The tSNE plots of CD86, MHCII, PDL1, and PDL2 producing myeloid cells (G) and percent frequencies of Mφ (H) and dendritic (I) cells producing these activation markers are shown. In bar graphs, individual data point and mean ± SEM values are plotted ( n = 5-mice/group). Significance (∗ p value ≤ 0.05) comparing two groups was analyzed by Student’s unpaired t test or Mann-Whitney U test and identified by a horizontal bar. Gating strategy and detailed data are available in and , respectively.

Article Snippet: FITC Rat anti-mouse anti-CD273 (PDL2) , ThermoFisher/eBioscience , RRID: AB_465462 ; 11-9972-82, 122, 488/520.

Techniques: Infection, Flow Cytometry, Expressing, Activation Assay, MANN-WHITNEY

Journal: Science Advances

Article Title: Ara-C suppresses H3 K27–altered spinal cord diffuse midline glioma growth and enhances immune checkpoint blockade sensitivity

doi: 10.1126/sciadv.adu3956

Figure Lengend Snippet: ( A ) qPCR was performed to analyze changes in immune checkpoint mRNA expression in PDCs with Ara-C for 0, 1, 3, and 5 days. n = 3 biological replicates per group. ( B ) Western blot showed the protein expression of different immune checkpoints in PDCs with Ara-C for 0, 1, 3, and 5 days. ( C ) Immunostaining showed the protein expression of different immune checkpoints in PDC_S01 with or without Ara-C treatment. ( D and E ) Representative images of flow cytometric results showed immune checkpoint expression on cell membranes of different PDCs. n = 3 biological replicates per group. ( F ) Representative images showed protein expression of immune-checkpoint ligands and receptors in tumors from C57 mice with or without treatment of Ara-C. n = 5 biological repeats. ( G ) Statistical comparison of immune checkpoint–positive cells between tumors with or without treatment of Ara-C. ( H ) Multiplex protein staining showed the expression of p21, PVR, and PD-L2 in spinal cord orthotopic xenograft tumors from C57 mice with or without treatment of Ara-C. ( I ) Multiplex protein staining showed the expression of CD3e, TIGIT, and PD-1 in spinal cord orthotopic xenograft tumors from C57 mice with or without treatment of Ara-C. ( J ) Multiplex protein staining showed the expression of CD3e, Granzyme B, and FOXP3 in spinal cord orthotopic xenograft tumors from C57 mice with or without treatment of Ara-C. *0.01 ≤ P < 0.05, **0.001 ≤ P < 0.01, ***0.0001 ≤ P < 0.001, and **** P < 0.0001 for all figures.

Article Snippet: The primary antibodies and dilutions used in this study were as follows: Histone H3 K27M Mutant Specific (1:1000, CST, 74829S), Tri-Methyl-Histone H3 Lys27 (1:200, CST, 9733S), Ki67 (ZSGB-BIO, ZM-0166), Cleaved Caspase-3 (1:400, CST, 9661S), p21 (1:1000, abcam, 188224), H3k9me3 (1:800, CST, 13969S), GFAP (1:300, abcam, ab68428), Nestin (1:200, abcam, ab105389), PDGFRA (1:500, abcam, ab203491), SOX2 (1:200, Proteintech, 11064), NeuN (1:500, abcam, ab177487), IBA1 (1:500, abcam, ab178846), PD-1 (1:1000, Proteintech, 66220-1), PDL2 (1:100, CST, 4918S), Galectin9 (1:300, abcam, ab69630), PVRL2 (1:500, Proteintech, 27171-1-AP), PVR (1:200, Proteintech, 83724-6), TIGIT (1:500, abcam, ab300073), CD3 (1:150, abcam, ab16669), and CD8 (1:1000, abcam, ab217344).

Techniques: Expressing, Western Blot, Immunostaining, Comparison, Multiplex Assay, Staining