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. 2019 Jan;29(1):83-86.
doi: 10.1038/s41422-018-0124-5. Epub 2018 Dec 4.

Palmitoylation stabilizes PD-L1 to promote breast tumor growth

Yi Yang  1 , Jung-Mao Hsu  1 , Linlin Sun  1   2 , Li-Chuan Chan  1   3 , Chia-Wei Li  1 , Jennifer L Hsu  1   3 , Yongkun Wei  1 , Weiya Xia  1 , Junwei Hou  1 , Yufan Qiu  1   4 , Mien-Chie Hung  5   6
Affiliations

Palmitoylation stabilizes PD-L1 to promote breast tumor growth

Yi Yang et al. Cell Res. 2019 Jan.
No abstract available

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Palmitoylation stabilizes PD-L1 to protect breast cancer cells from T-cell killing and promote tumor growth. a Western blot of PD-L1 in MDA-MB-231 (MB231) and BT549 cells treated with 2-BP as indicated for 12 h. b Western blot of PD-L1 in MB231 cells treated with 2-BP at 50 µM and harvested at the indicated time points. c PD-L1 palmitoylation was detected in MB231 and BT549 cells using streptavidin-HRP after ABE assay and immunoprecipitation with α-Flag beads. The same membranes were then re-probed with α-PD-L1 antibody. d PD-L1 palmitoylation was detected in primary human tumor tissues using streptavidin-HRP after ABE assay and immunoprecipitation with α-PD-L1 beads. One percent of tissue lysates served as input and were probed with α-PD-L1 antibody. e Detection of palmitoylation of PD-L1 and its C272A mutant in MB231 and BT549 cells (as MB231KO-PD-L1Flag, MB231KO-PD-L1C272A-Flag, BT549KO-PD-L1Flag, BT549KO-PD-L1C272A-Flag) using streptavidin-HRP after ABE assay and immunoprecipitation with α-Flag beads. The same membranes were then re-probed with α-PD-L1 antibody. f Western blot of PD-L1 in MB231KO-PD-L1Flag, MB231KO-PD-L1C272A-Flag and BT549KO-PD-L1Flag, BT549KO-PD-L1C272A-Flag cells treated with CHX and harvested at the indicated time points (left). The relative protein level of PD-L1 was quantified (right). Data represent mean ± SD of three independent experiments, ***P < 0.001, Student’s t test. g Detection of palmitoylation of mouse PD-L1 and its C272A mutant in 4T1 cells (as 4T1KO-mPD-L1Flag and 4T1KO-mPD-L1C272A-Flag) using streptavidin-HRP after ABE assay and immunoprecipitation with α-Flag beads. The same membranes were then re-probed with α-mPD-L1 antibody. h Western blot of mPD-L1 in 4T1KO-mPD-L1Flag and 4T1KO-mPD-L1C272A-Flag cells. i Top, T-cell-meditated tumor cell-killing assay in 4T1KO-mPD-L1Flag and 4T1KO-mPD-L1C272A-Flag cells along with mPD-L1 antibody or IgG treatment as indicated. Bottom, quantification of cell viability. Data represent mean ± SD of three independent experiments, ***P < 0.001, Student’s t test. j PD-L1Flag and ZDHHC9Myc expression plasmids were co-transfected into 293T cells, the interaction between PD-L1 and ZDHHC9 was determined by immunoprecipitation with α-Flag beads or α-Myc beads followed by immunoblotting with α-Myc or α-Flag antibody. One percent of whole cell lysates served as input. k PD-L1 palmitoylation was detected in MB231 and BT549 cells with ZDHHC9 knockdown as indicated using streptavidin-HRP after ABE assay and immunoprecipitation with α-PD-L1 beads. One percent of cell lysates served as input and were probed with α-PD-L1 antibody. l Western blot of PD-L1 in MB231 and BT549 cells with ZDHHC9 knockdown as indicated following CHX treatment at 100 µg/mL for 12 h. m mPD-L1 palmitoylation was detected in 4T1 and 4T1-ZDHHC9KO cells using streptavidin-HRP after ABE assay and immunoprecipitation with α-mPD-L1 beads. One percent of cell lysates served as input and were probed with α-mPD-L1 antibody. n Western blot of mPD-L1 in 4T1 and 4T1-ZDHHC9KO cells following INFγ treatment at 50 µg/mL for 12 h. o Top, T-cell-meditated tumor cell-killing assay in 4T1 and 4T1-ZDHHC9KO cells along with mPD-L1 antibody or IgG treatment as indicated. Bottom, quantification of cell viability. Data represent mean ± SD of three independent experiments, ***P < 0.001, Student’s t test. p Tumor growth of 4T1KO-mPD-L1Flag and 4T1KO-mPD-L1C272A-Flag cells in BALB/c mice. Tumors were measured at the indicated time points (n = 10 mice per group). q Tumor growth of 4T1 and 4T1-ZDHHC9KO cells in BALB/c mice treated with mPD-1 antibody or IgG as indicated. Tumors were measured at the indicated time points (n = 8 mice per group). Data represent mean ± SD. **P < 0.01, ***P < 0.001, Student’s t test
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