Journal: Nature Cell Biology

Article Title: SEL1L–HRD1 endoplasmic reticulum-associated degradation controls STING-mediated innate immunity by limiting the size of the activable STING pool

doi: 10.1038/s41556-023-01138-4

Figure Lengend Snippet: a , qPCR analysis in primary macrophages treated with Pam3 (TLR2) or LPS (TLR4) or transfected with double-stranded RNA (RIG-I) or cGAMP (STING) ( n = 4 mice each pooled from two independent repeats). mRNA, messenger RNA. b , ELISA analysis of secreted IFNβ and TNFα in primary macrophages treated with cGAMP for 6 h ( n = 3, 3, 7 and 8 mice (left to right), pooled from three independent repeats). c , Immunoblot analysis in primary macrophages transfected with vehicle (Veh) or cGAMP for 3 h, representative of four independent repeats. d , Immunoblot analysis in primary macrophages treated with vehicle or DMXAA for 1.5 h, representative of four independent repeats. e , ELISA analysis of secreted IFNβ and TNFα in primary macrophages transfected with poly(I:C) (RIG-1) for 18 h ( n = 3, 3, 4 and 4 mice (left to right); combined from two independent repeats). f , Immunoblot analysis in primary macrophages transfected with vehicle or poly(I:C) for 6 h, representative of two independent repeats. The relative intensity of p-STING (normalized to β-tubulin) or ratio of phosphorylated to total protein (p/t) are shown below the blots ( c , d and f ). All values represent means ± s.e.m. Statistical significance was determined by unpaired, two-tailed Student’s t -test ( a , b and e ).

Article Snippet: Inflammatory stimuli included the TLR2 ligand Pam3Cy (InvivoGen) at 1 μg ml −1 , the TLR4 ligand LPS (InvivoGen) at 500 ng ml −1 , the STING ligands 2′3′-cGAMP (InvivoGen), c-di-AMP (InvivoGen) and DMXAA (MedChemExpress) at 3.5 μg ml −1 , 3.5 μg ml −1 and 20 μg ml −1 , respectively, and the RIG-1 ligand poly(I:C) (InvivoGen) at 2 μg ml −1 .

Techniques: Transfection, Enzyme-linked Immunosorbent Assay, Western Blot, Two Tailed Test