Extended Data Fig. 5: Perturbation of STING activity in human LUAD models.
From: STING inhibits the reactivation of dormant metastasis in lung adenocarcinoma
(a) Western immunoblot analysis of STING and β-actin in WT and STING knockout H2087-LCC cells, representative of 2 independent experiments. (b–c) Immunoblotting (b) and qRT-PCR analyses (c) of STING expression in parental H2087-LCC cells and the spontaneous outbreak derivatives SO1 and SO2. Mean ± s.e.m, representative of 2 independent experiments. In (c), each dot represents a technical replicate of the assay. (d) qRT-PCR analysis of IFNB1 expression after cGAMP treatment of H2087-LCC, SO1, and SO2 cells. Mean ± s.e.m, representative of 2 independent experiments. Each dot represents a technical replicate of the assay. (e) Western immunoblot analysis of STING (endogenous STING and HA-tagged STING) and β-actin in H2087-LCC-SO cells inducibly expressing STING-HA under doxycycline (Dox) treatment for 24 h. Representative of 2 independent experiments. (f) Metastasis-free survival plots of athymic mice inoculated intracardially with 1 x 105 H2087-LCC-SO cells. Cells were transduced with a vector control or a Dox-inducible STING expression vector. Dox treatment started 7 days after cell inoculation. n = 13 (Control, STING), 10 (Control+Dox), 11 (STING+Dox) mice per group, log-rank test. (g) qRT-PCR analysis of STING mRNA levels in human primary bronchial epithelial cells, H2087, A549, and H2030 LUAD cell lines. Mean ± s.e.m, representative of 3 independent experiments. Each dot represents a technical replicate of the assay. (h–i) qRT-PCR analysis of IFNB1 mRNA expression in human primary bronchial epithelial cells and lung adenocarcinoma cell lines treated with cGAMP (h) or poly(I:C) (i). Mean ± s.e.m, representative of 3 independent experiments. Each dot represents a technical replicate of the assay. (j) BLI and quantification of metastases formed by H2030-BrM cells inducibly expressing activated STING-V155M, starting the day after intracardiac inoculation of 1 x 105 cells in athymic mice. Metastases were quantified 21 days after inoculation. n = 9 mice per group. Mean ± s.e.m., two-sided Mann-Whitney U-test. For gel source data, see Supplemental Fig. 1.
