Fig. 7. LD Induction following nucleic acid detection is EGFR mediated.

Primary immortalised astrocyte cells were treated with 2 μM AACOCF₃ (PLA₂ inhibitor) for 16 h prior to stimulation with dsRNA or dsDNA for 8 h and (a) were stained with Bodipy (493/503) to visualise LDs (green) and DAPI to visualise the cell nuclei (blue) 8 h post stimulation and (b) average numbers of LDs per cell was analysed using ImageJ analysis software (greater 200 cells, n = 2). c Primary immortalised astrocyte cells were treated with 2 μM AG-1478 (EGFR inhibitor) 16 h prior to stimulation with dsRNA or dsDNA, or treatment with OA and were stained with Bodipy (493/503) to visualise LDs and DAPI to visualise the cell nuclei 8 h post-stimulation. d The average number of LDs per cell analysed using ImageJ analysis software (greater 200 cells, n = 2). e Primary immortalised astrocyte cells were serum starved for 48 h, plated into wells and treated with 2 μM AG-14789 or control for 16 h. All cells were then given fresh full serum media for 36 h and stained to visualise LDs (green) as above. LDs were analysed using ImageJ analysis software. f Primary immortalised astrocyte cells were treated with 2 μM AG-1478 (EGFR inhibitor) for 16 h prior to stimulation with dsRNA and dsDNA for up to 72 h and were fixed at regular time points until 72 h post stimulation. Average numbers of LDs per cell was analysed using ImageJ analysis software (greater than 200 cells, n = 2). g Primary immortalised astrocyte cells were treated with 2 μM AG-1478 (EGFR inhibitor) for 16 h prior to stimulation with IFN- β and their LDs were numbers assessed using image J analysis software. In (b–g) error bars, mean values ± SEM, P values were determined by unpaired two-tailed Student’s t test with a Holm-Sidak correction for multiple comparisons for 2 or more groups (greater than 300 cells; n = 3 biological replicates). Stimulated cells were statistically compared with their respective mock controls, ns = not significant. h Primary immortalised astrocyte cells were treated with 2 μM AACOCF₃ for 16 h prior to stimulation with dsRNA or dsDNA for 16 h where secreted IFN protein levels from these experiments were analysed via ELISA for IFN- β and IFN-λ protein. Error bars, mean values ± SEM, P values were determined by two-way ANOVA post-hoc pairwise comparisons with Bonferroni correction (n = 3 biological replicates). Source data are provided as a Source Data file.