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. 2021 Jul 14;14:657064. doi: 10.3389/fnmol.2021.657064

FIGURE 1.

FIGURE 1

Experimental design and workflow. (A) Male and female mice were treated with nicotine or control solutions. In C3H/HeJ mice, sub-chronic nicotine or saline was administered subcutaneously on alternating days for a total of 6 days in a CPP testing paradigm (n = 4 per group). In C567BL/6J mice, nicotine or saccharin was administered in drinking water chronically for 21 days, followed by a 24-h withdrawal period (n = 4 per group). About 10–30 min after CPP testing in C3H/HeJ mice, or 24 h after cessation of nicotine or saccharin treatment in C57BL/6J mice, subjects were sacrificed and the VTA and NAc shell were collected from each brain by 1 mm punch biopsy on 1 mm-thick coronal sections. (B) Tissue samples were subsequently processed for proteomic analysis with a TMT10-plex isobaric labeling strategy. (C) Data were quantified, normalized, analyzed, and visualized using MaxQuant and Perseus software and the online STRING database tool.