Fig. 4
From: Genetically encoded phosphatidylserine biosensor for in vitro, ex vivo and in vivo labelling
The expression of C2 fusion proteins in ex vivo organotypic hippocampal slices after AAV delivery. a OHSC were transduced with AAV carrying either C2-SNAP or C2m2-SNAP at 5 days in vitro (5DIV) and maintained for 9 days for protein expression. The secretion of recombinant proteins was inhibited by 10 µg/ml brefeldin A (BFA) for 5 h. Fixed control (CTRL) and inhibited (BFA) slices were labelled by immunohistochemistry and SNAP-tag substrate for fluorescent imaging. Created by Biorender.com. b Confocal images of OHSC transduced with either AAV-C2-SNAP or AAV-C2m2-SNAP. Cell nuclei were stained with DAPI, fused SNAP-tag was labelled with AlexaFluor647 (AF647). Astrocytes, microglia and neurons were labelled with GFAP, Iba1 antibodies, or expressed EGFP, respectively. Scale bar 20 µm. c, d Quantification of C2-SNAP and C2m2-SNAP fluorescence within astrocytes with or without BFA treatment. Data presented as means ± standard error of the mean (n = 10–15 images per replicate, two independent replicates). Means were compared by one-way ANOVA and post-hoc Tukey test. p values < 0.05 were considered as significant. RFU relative fluorescence units