Fig 2: Neither Akt2 nor Akt3 compensates for Akt1-/- at the mRNA or protein level at PND50. A) Representative Western blots of Akt1+/+, Akt1+/-, and Akt1-/- ovaries at PND50. Shown are phosphorylated Ser473 AKT and Thr308 AKT1, and AKT1, AKT2, and AKT3 total protein; ß-actin was used as a loading control. Changes in the relative mRNA expression of Akt2 (B) and of Akt3 (C) in Akt1-/- (gray bars) ovaries relative to Akt1+/+ (black bars) control ovaries. Expression was determined by using the Pfaffl method.

Fig 3: MNRR1 upstream of known metastasis-associated genes. a List of genes identified by RNA-Seq to be significantly downregulated in the HEK293 R1-KO cells and rescued in the R1-KO cells expressing the transcriptionally active version of MNRR1 (R1-TA). The PubMed IDs (PMID) of the studies published indicating the role of each of the genes in cancer metastasis is shown. b AKT3 (circled) is the most upregulated gene in the MNRR1-knockout cells. c Total AKT3 protein levels in WT, R1-KO, and R1-OE cells with Actin used as a loading control. d MCF7 cells (WT) with a knockout of MNRR1 (MCF7 R1-KOm; KO) also display an increase in total AKT3 protein levels. GAPDH is used as a loading control

Fig 4: IFN-? induces LRRK2 expression in human iPSC-derived neurons and decreases AKT phosphorylation.IFN-? treatment induces an increase of LRRK2 levels in human iPSC-derived neurons. a LRRK2 mRNA levels in control iPSC-derived neurons treated with IFN-? 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-? at the indicated concentrations (mean ± SEM, one-way ANOVA, Bonferroni post hoc, *P < 0.05; n = 3 independent experiments). c IFN-? transcriptional signature genes in control iPSC-derived neurons treated with 200 IU/mL IFN-? 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-?, expressed as fold changes relative to corresponding IFN-? 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-?, expressed as fold changes relative to corresponding IFN-?-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-? is indicated (n = 3 independent experiments). g Quantification of phospho-AKT3/AKT3 ratio in LRRK2 G2019S neurons and isogenic controls, untreated and upon IFN-? 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).

Fig 5: Downregulation of Akt3 mimics the effects of miR-207. (a) Expression levels of Akt3 and other Akt isoforms, such as Akt1 and Akt2, were determined using western blotting analyses in HEI-OC1 cells transfected with siAkt3 or control. (b) HEI-OC1 cells were transfected with miR-207, siAKT3 or control. The apoptosis assay using flow cytometry was performed 24 h after transfected cells were treated with 20 Gy IR. Error bars, S.D.; *P=0.001 (n=3) versus control. (c) Expression of ?-H2AX, cleaved PARP and Akt3 proteins were determined by western blotting analyses using the indicated antibodies in transfected cells after 20 Gy IR. The cells used to determine the expression level of ?-H2AX and cleaved PARP and Akt3 were collected at 12 h and 24 h after IR, respectively