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. 2023 Mar 28;73(1):131–155. doi: 10.1136/gutjnl-2022-327927

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© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY. Published by BMJ.

This is an open access article distributed in accordance with the Creative Commons Attribution 4.0 Unported (CC BY 4.0) license, which permits others to copy, redistribute, remix, transform and build upon this work for any purpose, provided the original work is properly cited, a link to the licence is given, and indication of whether changes were made. See: https://creativecommons.org/licenses/by/4.0/.

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Co-depletion of FAK and STAT3 potentiates antitumour immunity. (A) Fold change in tumour weight relative to FAK-/- tumours 2 weeks postimplantation into the pancreas of C57BL/6 mice. n=6 tumours per group. (B) Fold-change in tumour weight relative to FAK-/- tumours 4 weeks postimplantation into the pancreas of C57BL/6 mice. n=6 tumours per group. (C) Left, flow cytometry quantification of MHC-I-expressing cells as a percentage of CD45^- cells from FAK-/- and FAK-/-STAT3shRNA tumours. Right, flow cytometry quantification of the median fluorescence intensity of MHC-I expression in CD45^- cells from FAK-/- and FAK-/-STAT3shRNA tumours. n=5 tumours per group. (D) Flow cytometry quantification of CD45⁺ cells as a percentage of live cells from FAK-/- and FAK-/-STAT3shRNA tumours. n=5 tumours per group. (E) Flow cytometry quantification of effCD8⁺ T-cells represented as number of cells per mg of tumour tissue (left) and as a percentage of CD45⁺ cells (right) from FAK-/- and FAK-/-STAT3shRNA tumours. n=5 tumours per group. (F) Representative images of FAK-/- and FAK-/- STAT3shRNA tumour sections stained with anti-CD8 or anti-granzyme-B antibodies. (G) Flow cytometry quantification of PD-1⁺ cells as a percentage of effCD8⁺ T-cells from FAK-/- and FAK-/-STAT3shRNA tumours. n=5 tumours per group. (H) Fold change in tumour weight relative to FAK-wt tumours 2 weeks postimplantation into the pancreas of C57BL/6 mice. n=6 tumours per group. (I) Representative images of FAK-wt and FAK-wt STAT3shRNA tumour sections stained with anti-CD8 or anti-granzyme-B antibodies. (J) Quantification of the number of CD8 T-cells per field of view (FOV) from F and I. (K) Quantification of the relative area positive for Granzyme B staining per FOV from (F, I). (L) Proteome profiler array analysis of chemokine secretion from FAK-wt, FAK-/-, FAK-wt STAT3shRNA and FAK-/- STAT3shRNA cells±10 ng/mL IFNγ stimulation for 24 hours. Data in (A–E, G, H, J, K) represented as mean±SEM. Statistical significance in (A–E, G and H) calculated using a two-tailed unpaired t-test. Statistical significance in J and K calculated using a Kruskal-Wallis test with Dunn’s multiple comparison. *p≤0.05, **p≤0.01, ***p≤0.001, ****p≤0.0001. FAK, focal adhesion kinase; MHC-I, Major Histocompatibility Complex class-I.