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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: Sirtuin 5 aggravates microglia-induced neuroinflammation following ischaemic stroke by modulating the desuccinylation of Annexin-A1

Fig. 4

SIRT5 regulates the subcellular localization of ANXA1. A Primary cultured microglia were infected with adenovirus encoding HA-ANXA1-WT, HA-ANXA1-166R or HA-ANXA1-166E and subjected to OGD/R. Western blot assay indicating ANXA1 levels in membrane and nuclear fractions. B Immunofluorescence assays of HA-ANXA1 (red) with colabelled DAPI (blue, nuclei), showing the subcellular location of HA-ANXA1. C Quantitative analysis of nuclear/cytoplasmic HA-ANXA1 levels in (B). D Primary cultured microglia were infected with adenoviruses encoding Flag-SIRT5-WT and Flag-SIRT5-H158Y and subjected to OGD/R. Western blot assay indicating ANXA1 levels in membrane and nuclear fractions. E Immunofluorescence assays of ANXA1 (red) with colabelled DAPI (blue, nuclei), showing the subcellular location of ANXA1. F Quantitative analysis of nuclear/cytoplasmic ANXA1 levels in (E). G Primary cultured microglia were infected with adenovirus encoding the SIRT5 shRNA sequence and subjected to OGD/R. Western blot assay indicating ANXA1 levels in membrane and nuclear fractions. H Immunofluorescence assays of ANXA1 (red) with colabelled DAPI (blue, nuclei), showing the subcellular location of ANXA1. I Quantitative analysis of nuclear/cytoplasmic ANXA1 levels in (H). Scale bars, 20 μm. Data are presented as the mean ± SEM from at least three independent experiments. ns: no significance, **P < 0.01, ***P < 0.001, and ****P < 0.0001

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