2.1 Animals
Two types of C57BL/6J mice, Cnr1flox/flox (non-cKO) and PV-Cre;Cnr1flox/flox (Cnr1-cKO or
PV;Cnr1cKO) were used for the experiments. Mice were
bred and housed according to the guidelines of Institutional Animal Care
and Use Committee of Anhui University (protocol numbers 2020-039).
Genotyping of the transgenic mice was performed using PCR and agarose
gel electrophoresis with the primers listed in Table S1 to identify
homozygous and hemizygous mice. The mice were housed in a facility with
controlled temperature and humidity on a 12-hour light/dark cycle
(lights on at 8 a.m.), and were provided with food and water ad libitum
until they reached 8–10 weeks of age. In the immunostaining analysis
for CB1Rs distribution, the number of non-cKO male mice used for each
auditory nucleus was as follows: CN (n=10), SOC (n=7), LL (n=5), IC
(n=8), AC (n=3), and MGB (n=3) (Fig. 1, Fig. 2). For the double-staining
analysis of PV and CB1Rs, both non-cKO (n=5) and Cnr1-cKO (n=5) mice
were used (Fig. 3). In the western blot and RT-qPCR analyses, the number
of mice used were as follows: wild-type (n=5), Cnr1-cKO (n=5), and
non-cKO (n=6) (Fig. S6). For the ABR tests, non-cKO (n=5) and Cnr1-cKO
(n=5) mice were used (Fig. 4).