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. 2020 Sep 25;11:4855. doi: 10.1038/s41467-020-18634-y

Fig. 4. ACKR3 stimulation regulates Cx43 cellular localization and inhibits GJIC in astrocytes.

Fig. 4

a Confocal images of Cx43 immunostaining of confluent primary astrocyte cultures from WT or β-arr2−/− mice representative of three independent experiments performed on different sets of cultured cells exposed for 30 min to either vehicle or CXCL12 (10 nM) or CXCL11 (100 nM) in the absence or presence of Dynasore (Dyn, 80 μM). Scale bar = 50 μm. The histograms show the size of Cx43 gap junctional plaques (clusters of channels present at the cell-cell interface). Data were normalized to the one measured in vehicle-treated cultures. They are the means ± SEM of values (Two-way ANOVA, Bonferroni post-hoc F(4,18)). b Representative photomicrographs of scrape loading in confluent primary astrocyte cultures obtained from embryonic WT and β-arr2−/− mice taken 10 min after the scrape in the presence of LY. CXCL12 (10 nM), CXCL11 (100 nM), and Dynasore (Dyn, 80 μM) were applied for 30 min. CBX (50 μM) was applied overnight. Scale bar = 200 μm. LY diffusion was calculated by measuring the distance from the scrape where LY fluorescence intensity is 50% of the maximal fluorescence. Values were normalized to LY diffusion in vehicle-treated astrocytes. Results represent the means ± SEM of values (Two-way ANOVA, Bonferroni post-hoc, F(6,24)). See the Statistics and Reproducibility section for the number of repetitions, exact P values and symbol (*/$) legend. Source data are provided as a Source Data file.