Fig. 1: IFN-γ induces LRRK2 expression in human iPSC-derived neurons and decreases AKT phosphorylation.

IFN-γ treatment induces an increase of LRRK2 levels in human iPSC-derived neurons. a LRRK2 mRNA levels in control iPSC-derived neurons treated with IFN-γ for 24 h at the indicated concentrations. LRRK2 mRNA levels, measured by qRT-PCR, are expressed as fold changes relative to untreated (mean ± SEM, one-way ANOVA, Bonferroni post hoc, ****P = 0.0001, ***P = 0.0003, **P = 0.0011, n = 3 independent experiments). b Representative Western blots and corresponding quantification of LRRK2 expression in control (left) and isogenic LRRK2 G2019S (right panel) iPSC-derived neurons treated with IFN-γ at the indicated concentrations (mean ± SEM, one-way ANOVA, Bonferroni post hoc, *P < 0.05; n = 3 independent experiments). c IFN-γ transcriptional signature genes in control iPSC-derived neurons treated with 200 IU/mL IFN-γ for 24 h measured by qRT-PCR and expressed as fold changes relative to untreated controls. Downregulated (left panel) and moderately (fold change <5, middle panel) or strongly (fold change >5, right panel) upregulated genes are shown (mean ± SEM, two-tailed t-test treated vs untreated, left panel: **P = 0.0075, 0.0075, 0.0052 in sequence, middle panel: *P = 0.0186, 0.0379 in sequence, right panel: ****P = 0.0001, ***P = 0.0003, **P = 0.0080, *P = 0.0356, 0.0238 in sequence; n = 3 independent experiments). d mRNA levels of AKT3, STAT1, SOCS1, ICAM1, and IRF1 in LRRK2 G2019S (G2019S-1) neurons treated with 200 IU/mL IFN-γ, expressed as fold changes relative to corresponding IFN-γ treated isogenic controls (CTRL-1) (mean ± SEM, two-tailed t-test, **P = 0.0060; n = 3 independent experiments). e mRNA levels of AKT3 in LRRK2 G2019S neurons (G2019S-2, G2019S-3) treated with 200 IU/mL IFN-γ, expressed as fold changes relative to corresponding IFN-γ-treated isogenic controls (CTRL-2, CTRL-3) (mean ± SEM, two-tailed t-test, **P = 0.0088, 0.0033 in sequence; n = 3 independent experiments). f Representative Western blot of AKT3 in LRRK2 G2019S neurons (G2019S-1, G2019S-2) and corresponding isogenic controls (CTRL-1, CTRL-2). Treatment with 200 IU/mL IFN-γ is indicated (n = 3 independent experiments). g Quantification of phospho-AKT3/AKT3 ratio in LRRK2 G2019S neurons and isogenic controls, untreated and upon IFN-γ treatment, as assessed by Luminex multiplex analysis (mean ± SEM, two-way ANOVA, Bonferroni post hoc, **P = 0.0033, 0.0021 in sequence, *P = 0.0270, 0.0165 in sequence; n = 4 independent experiments).