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. 2019 Apr 18;216(6):1359–1376. doi: 10.1084/jem.20180660

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© 2019 Medina et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 2. — Batf3 dependence distinguishes resident memory from antitumor effector CD8⁺ T cells. (A) Flow cytometry and frequency of PD-1 and KLRG1 on CD8⁺ T cells from tumors of untreated Kit^V558Δ/+;Batf3^+/+ and Kit^V558Δ/+;Batf3⁻^/⁻ mice (six mice/group pooled from two experiments). (B) Tumor cell suspensions from Kit^V558Δ/+;Batf3^+/+ and Kit^V558Δ/+;Batf3⁻^/⁻ mice were stimulated with PMA and ionomycin in the presence of brefeldin A followed by intracellular analysis of IFN-γ and TNF in CD8⁺ T cells (five mice/group, representative of two independent experiments). (C) Normalized gene expression of BATF3 was plotted against CD8B, IFNG, and the cytolytic score from 26 untreated human GIST specimens. (D) Untreated tumors in Kit^V558Δ/+;Batf3^+/+ and Kit^V558Δ/+;Batf3⁻^/⁻ mice were stained for CD8. Top bar represents 200 µm. Black arrowheads point to staining in periphery and correspond to top inset. White arrowheads point to staining in central core and correspond to lower inset. Inset bar represents 50 µm (representative of three mice/group). (E) Flow cytometry and frequency of CD103 and CD69 on CD8⁺ T cells from untreated tumors of Kit^V558Δ/+;Batf3^+/+ and Kit^V558Δ/+;Batf3⁻^/⁻ mice (six mice/group pooled from two experiments). (F) Flow cytometry and frequency of PD-1 on CD103⁺ resident and CD103⁻ effector CD8⁺ T cells and PD-1 MFI of PD-1⁺ effector and PD-1⁺ resident CD8⁺ T cells from tumors of Kit^V558Δ/+;Batf3^+/+ mice (five mice/group, representative of two independent experiments). (G) Flow cytometry and frequency of CD44 and CD62L on resident CD8⁺CD103⁺ and effector CD8⁺CD103⁻ T cells from tumors of Kit^V558Δ/+;Batf3^+/+ and Kit^V558Δ/+;Batf3⁻^/⁻ mice (five mice/group, representative of two independent experiments). (H) Flow cytometry and frequency of CD44 and Tbet on resident and effector CD8⁺ T cells from tumors of Kit^V558Δ/+;Batf3^+/+ and Kit^V558Δ/+;Batf3⁻^/⁻ mice (four mice/group, pooled from two experiments). Data represent mean ± SEM; P values were calculated using a Student’s t test; *, P < 0.05. Correlation is shown using R², and significance was determined using Spearman correlation.