Fig. 4: UBE3A is highly enriched in the cytoplasm of macrophages.

A–C Bone marrow-derived macrophages (BMDMs) were treated with myelin (50 µg/ml) for 24 h (Mye²⁴), 72 h (Mye⁷²), or left untreated (Ctrl). Representative images of UBE3A and BODIPY (A). Quantification of the number of UBE3A⁺ puncta in the cytoplasm and the nucleus (B) and the ratio of UBE3A puncta in cytoplasm:nucleus (C) (n = 3 samples from three independent experiments, >20 cells per condition). Scale bar 50 µm. D Upper figure shows a schematic representation of the number of Ube3a exon junctions found in BMDMs. Two lower schemes depict splicing pattern and relative abundance of predicted Ube3a transcripts in untreated macrophages, based on quantification of exon junctions (n = 3 samples). E BMDMs were treated with myelin (50 µg/ml) for 24 h (Mye²⁴), 72 h (Mye⁷²), or left untreated (Ctrl). Cell lysates were analyzed by immunoblotting for UBE3A, normalized to GAPDH levels (loading control), and the relative isoform abundance ratio was determined. Upper UBE3A band is mIso2, lower UBE3A band is mIso3 (n = 6 samples from six independent experiments). Data were represented as mean ± SEM. Statistical significance was determined by two-sided one-way ANOVA (B, C, E). *P < 0.05 and ***P < 0.001. If p values are not indicated, the result was not significant.