Fig. 1

Characterization of an in vitro model of neuroinflammation and expression of α7 nAchRs in mouse astrocytes. a These are representative images of mouse astrocytes using the Cellomics ArrayScan XTI high-content analysis system for automated image acquisition of 20 fields for each well at × 20 magnification. In each field, two channels were captured to image nuclei (DAPI) and astrocyte marker (GFAP). The ArrayScan Target Activation bioapplication was used for processing and analyzing the images to quantify GFAP positive cells as percentage of total cell counts using DAPI within a well. This was found to be more than 90%. b LPS (60 ng/ml) treatment for 3 h increased nuclear translocation of p65 subunit of NF-κB in astrocytes as a marker of acute inflammation phase. c LPS (60 ng/ml) treatment for 24 h resulted in significant increase in pro-inflammatory cytokines, TNF and IL-6, downstream of NF-κB activation in astrocytes, *p < 0.05 as compared with untreated (t test). Error bars represent SD (n = 4). d Stimulation of astrocytes with LPS (60 ng/ml) for 24 h increased cell processes substantially as compared to the control cells. e α7 nAChR subunit mRNA expression observed in cultured astrocytes (three replicates). f Binding of fluorescently labeled α-bungarotoxin was observed in astrocytes. Pretreatment with 200 μm nicotine (α7 nAchR agonist), significantly inhibited binding of fluorescently labeled α-bungarotoxin shown by decrease in fluorescence confirming expression of α7 nAchR in these cells