ab210073 anti cd45 imc cell signaling technology  (Cell Signaling Technology Inc)

Journal: Cancer cell

Article Title: Translation dysregulation in cancer as a source for targetable antigens.

doi: 10.1016/j.ccell.2025.03.003

Figure Lengend Snippet: Figure 6. Longitudinal scRNA-seq analysis of tumor immune compartment reveals anti-tumor CD8+ T cell responses linked to Tyw2 KO (A) In vivo tumor growth following inoculation of WT19, KO13, or KO18 cells into C57BL/6 mice. Time points highlighted with dotted box were used for scRNA-seq analysis. ***p < 0.001, pairwise Bonferroni-corrected Student’s t test. (B) Schematic overview of experimental workflow. Tumors were removed and dissociated at day 18 and day 21 prior to MULTI-seq barcoding, enrichment for CD45+ immune cells, and scRNA-seq. (C) Uniform manifold approximation and projection (UMAP) of immune gene expression space colored by cell type. (D) Bar charts of the percentage of CD8+ T cells among all CD45+ cells in each tumor background at day 18 (D18) and day 21 (D21). Statistically significant changes denoted with propeller test p values (ns, not significant; n = 3 sets of 4–5 pooled tumors). (E) UMAP of CD8+ T cell gene expression space colored by subtype. (F) Dot plot of CD8+ T cell subtype annotation genes. Dot color indicates expression level and size indicates the proportion of cells expressing each gene. (G) UMAP of CD8+ T cell gene expression space colored by sample identities on day 18 (top) and day 21 (bottom). Regions of gene expression space that are enriched in Tyw2 KO or WT samples highlighted with dotted circles. (H) Bar charts of the percentage of exhausted and memory-like CD8+ T cells among all CD8+ T cells in each tumor background at day 18 and day 21. Statistically significant changes denoted with propeller test p values (ns, not significant; n = 3 sets of 5 pooled tumors). (I) Z score heatmap of the average expression of immunosuppression markers in CD8+ T cell subtypes. Z scores for each gene clustered using hierarchical clustering. Exhausted CD8+ T cells highlighted with red box. (J) Representative images of the day 21 Tyw2 KO and WT TME with bar charts of mean ± SEM total (top), exhausted (middle), and proliferative CD8+ T cell percentages (bottom). Exact p values from Welch two-sample t test (n = 5 mice per tumor background). (K) Weighted network graph of IFNG-IFNGR1/IFNGR2 predicted signaling interactions split by tumor background and time point. Nodes colored by cell type, edges weighted by signaling probability and colored by sender cell type. See also Figure S6.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies anti-TYW2 (TRMT12) Novus-Biologicals Cat#NBP1-76583; RRID: AB_11025190 anti-a tubulin mouse Millipore Cat#05-829; RRID: AB_310035 anti-mCherry Abcam Cat#Ab167453; RRID: AB_2571870 anti-rabbit HRP conjugate Cell Signaling Technology Cat#5127; RRID: AB_10892860 anti-mouse HRP conjugate Cell Signaling Technology Cat#91196; RRID: AB_2940774 pan-HLA antibody W6/32 ATCC ATCC-HB-95RRID; N/A APC anti human HLA A,B,C BioLegend Cat# 311410; RRID: AB_314879 PE anti-human CD3 BioLegend Cat#300308; RRID: AB_314044 FITC anti-human CD8 BioLegend Cat#301060; RRID: AB_2564165 APC anti-human CD137 (41BB) BioLegend Cat#309810; RRID: AB_830672 APC anti- human IFNg BioLegend Cat#502512; RRID: AB_315237 APC anti-human TNFa BioLegend Cat#502912; RRID: AB_315264 APC anti-human CD8 BioLegend Cat#300912; RRID: AB_314116 PE-Cy7 anti-human CD3 BioLegend Cat# 300316; RRID: AB_314052 PE anti- human IFNg BioLegend Cat#502509; RRID: AB_315234 BV421 anti-human TNFa BioLegend Cat# 502932; RRID: AB_10960738 InVivoMAb anti-mouse CD8a, clone 2.43 BioXcell Cat#BE0061; RRID: AB_1125541 InVivoMAb rat IgG2b isotype control, clone LTF-2 BioXcell Cat#BE0090; RRID: AB_1107780 InVivoMAb anti-mouse PD-1 (CD279), clone RMP1-14 BioXcell Cat#BE0146; RRID: AB_10949053 InVivoMAb rat IgG2a isotype control, clone 2A3 BioXcell Cat#BE0089; RRID: AB_1107769 CD8a (D4W2Z) XP Rabbit mAb Cell Signaling Technology Cat#60168RRID; N/A Anti-CD3 epsilon antibody [CAL57] abcam Ab251607RRID; N/A Anti-LAG-3 antibody [CAL77] abcam Ab251606RRID; N/A CD45 (D3F8Q) Rabbit mAb Cell Signaling Technology Cat#98819RRID; N/A BD Pharmingen Purified Mouse Anti-Ki-67 BD Biosciences Cat#556003; RRID: AB_396287 FITC anti-mouse CD3 BioLegend Cat#100203; RRID: AB_312660 (Continued on next page) Cancer Cell 43, 1–18.e1–e18, May 12, 2025 e1

Techniques: In Vivo, Gene Expression, Expressing