Extended Data Fig. 6: STING activity in mouse lung adenocarcinoma cells.
From: STING inhibits the reactivation of dormant metastasis in lung adenocarcinoma
(a-b) qRT-PCR analysis of Sting (a) and Cxcl10 (b) mRNA levels after cGAMP treatment in KPad1 and KP-482T1 cells. Mean ± s.e.m, representative of 2 independent experiments. Each dot represents a technical replicate of the assay. (c) Western immunoblotting of STING and β-actin in parental and Sting knockout KPad1 cells. Representative of 2 independent experiments. For gel source data, see Supplemental Fig. 1. (d) RFP-positive WT KPad1 cells and GFP-positive Sting KO KPad1 cells were mixed at the indicated ratios and intracardially inoculated (1 x 105 cells) in B6-albino mice. Metastases-bearing organs were harvested 4-17 weeks after inoculation and the percentage of GFP-positive cells and RFP-positive cells was determined by flow cytometry. n = 9 (1:1, 9:1) or 5 (3:1) lesions from 3-5 mice per group. Mean ± s.e.m. (e) Percentage of EdU positive cells of WT or Sting knockout KPad1 cells incubated with EdU for 2 h. n = 3 per group. Mean ± s.e.m. (f) Percentage of EdU positive cells in KP-482T1 Tet-On Sting cultures treated with Dox for 24 h and then incubated with EdU for 2 h. n = 3 per group. Mean ± s.e.m. (g–h) BLI signal intensity at the indicated time points after intracardiac inoculation of 1 x 105 WT or Sting knockout KPad1 cells (g) or KP-482T1 cells with induced overexpression of STING (h) in B6-albino mice. n = 16 (WT) or 15 (KO) mice (g); n = 7 (control) or 6 (Dox) mice (h). Mean ± s.e.m. (i) Heatmap representation of cytokine array analysis of culture supernatant from KP-482T1 cells transduced with a vector control or a Sting overexpression vector. n = 3 for each condition.
