A total of 9 female C57BL/6 mice (seven weeks old) were obtained from Beijing Vital River Laboratory Animal Technology Co. Ltd. They were randomly divided into three groups (n = 3 per group). One group served as the control, another group underwent ovarian removal, and a third group underwent ovarian removal and received E2. Ovariectomy was performed following the methods described in another study [36 (link)]. Ovariectomized (OVX) mice were intraperitoneally injected with PBS or E2 (100 µg/kg) every day. The LLC cells (1 × 106) expressing firefly luciferase were injected directly into the left lung (in 50 µL of 1:1 mix of PBS and Matrigel (BD Biosciences)) three days after surgery [37 ]. After 12 days, bioluminescence imaging experiments were performed using a Bruker In-Vivo MS FX Pro small animal optical imaging system.
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