Figure 2 | Scientific Reports

Figure 2

From: PP2A phosphatase inhibition is anti-fibrotic through Ser77 phosphorylation-mediated ARNT/ARNT homodimer formation

Figure 2

Serine77 phosphorylation site of ARNT is responsible for the homodimer formation. (a) ARNT phosphorylation site is identified at the serine on position 4 of the peptide (Ser77 in ARNT) after 5 µM of LB100 treatment. (b) Domain organization of ARNT (NLS-nuclear localization domain; bHLH-basic helix loop helix; PAS-periodicity/ARNT/single-minded; TAD-transactivation domain). (c) EGFP or myc-tagged wild type ARNT overexpression constructs in which serine 77 was changed to the aspartic acid residue. (d, e) Representative immunoblotting for co-immunoprecipitation of EGFP or myc-tagged wildtype and mutated ARNT expression construct after 4 h of LB100 treatment (5 µM). (f) Representative confocal images of proximity ligation assay (PLA) for EGFP and myc-tagged wild type and mutated ARNT expressing HEK293 cells after LB100 (5 µM) treatment. Scale bars: 15 µm. (g) Quantitative analysis showing PLA signals, measured as dots per cells (N = 20 cells). ****p < 0.0001, 1-way ANOVA versus IgG. Data are shown as mean ± SD.

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