Fig. 2: LRRK2 inhibits nuclear NFAT3 shuttling in human neurons. | Nature Communications

Fig. 2: LRRK2 inhibits nuclear NFAT3 shuttling in human neurons.

From: Interferon-γ signaling synergizes with LRRK2 in neurons and microglia derived from human induced pluripotent stem cells

Fig. 2: LRRK2 inhibits nuclear NFAT3 shuttling in human neurons.

a Representative Western blots of NFAT3 in nuclear (N) and cytosolic (C) fractions of LRRK2 G2019S neurons and isogenic controls (left and middle panel). Treatment with 200 IU/mL IFN-γ is indicated. On the right, quantification of nuclear to total (N and C fraction) NFAT3 in LRRK2 G2019S iPSC-derived neurons and isogenic controls (mean ± SEM, two-way ANOVA, Bonferroni post hoc, ***P = 0.0003, **P = 0.0015, 0.0035 in sequence, *P = 0.0226; n = 5 independent experiments). b Representative Western blots of NFAT3 in N and C fractions of control (left panel) and isogenic LRRK2 G2019S neurons (middle panel). Treatments with 1 μM ionomycin for 30 min and 200 IU/mL IFN-γ for 24 h are indicated. On the right, quantification of nuclear to total (N and C fraction) NFAT3 in LRRK2 G2019S iPSC-derived neurons and isogenic controls (mean ± SEM, one-way ANOVA, Bonferroni post hoc, ****P < 0.0001, ***P = 0.0005, 0.0008, 0.0002 in sequence; n = 5 independent experiments). c Representative Western blot (left) and quantification (right) of NFAT3 from whole-cell extracts in LRRK2 G2019S neurons and isogenic controls. Treatments with 200 IU/mL IFN-γ are indicated (mean ± SEM, two-way ANOVA, Bonferroni post hoc, ***P = 0.0009, *P = 0.0421; n = 5 independent experiments). d Representative Western blot (left) and quantification (right) of NFAT3 from whole-cell extracts in control iPSC-derived neurons upon treatment with 200 IU/mL IFN-γ for 24 h, with and without treatment with a proteasome inhibitor (MG132, 20 nM for 24 h) (mean ± SEM, one-way ANOVA, Bonferroni post hoc, *P = 0.0399, 0.0437 in sequence; n = 3 independent experiments). e Representative Western blots of NFAT3 in nuclear and cytosolic fractions of LRRK2 KO neurons. Treatments with 1 μM ionomycin for 30 min and 200 IU/mL IFN-γ for 24 h are indicated. On the right, quantification of nuclear to total (N and C fraction) NFAT3 (mean ± SEM, one-way ANOVA, Bonferroni post hoc, ****P < 0.0001; n = 5 independent experiments).

Back to article page