Extended Data Fig. 8: Analysis of Acly+Ep300+ astrocytes by FIND-seq.
From: Disease-associated astrocyte epigenetic memory promotes CNS pathology
(a) FACS sorting schematic for tdTomatoGfap astrocytes. (b-d) Schematic illustration of microfluidic devices utilized in FIND-seq. (b) The bubble-triggered device has with five inlets: i) cell inlet, ii) Oligo dT primer-conjugated agarose inlet, iii) lysis buffer inlet, iv) oil inlet, and v) pressurized air inlet. (c) The re-injector device has three inlets: i) agarose bead inlet, ii) TaqMan PCR master mix inlet, and iii) oil inlet. (d) The droplet sorter device has seven inlets: i) emulsion inlet, ii) spacing oil inlet, iii) additional spacing oil inlet, iv) bias oil inlet, v) saltwater inlet (for the electrode), vi) saltwater inlet (for the moat), and vii) pressurized air inlet. (e) Astrocytes are encapsulated in an agarose bead along with the lysis buffer. The genome entrapped in the agarose bead is stained with SYBR Green I and visualized using a fluorescence microscope. (f) cDNA, produced on the agarose bead, is amplified via WTA and validated using a Bioanalyzer. (g) Principal component analysis (PCA) plot of Acly−Ep300−, Acly+Ep300−, Acly−Ep300+, Acly+Ep300+ EAE astrocytes. (h) Violin plot depicting Acly+Ep300+ EAE astrocyte signature expression in EAE astrocytes. (i) IPA pathway analysis up-regulated (red) in cluster 1 astrocytes. (j) EP300-NF-κB signaling network of cluster 1 astrocytes. (k) IPA pathway analysis up-regulated (red) in cluster 10 astrocytes. (l) EP300-NF-κB signaling network of cluster 10 astrocytes.
