(A) Scheme of the Cre inducible shRNA expression system used to selectively
silence p11 expression. (B) HEK 293 cells were transfected with the indicated constructs
and probed for p11 expression 72hrs later. In the presence of Cre lox.sh.p11 construct
silences p11 with a similar efficiency than the constitutively active sh.p11. (C) D1-Cre
mice injected in the NAc with Cre inducible AAV2-lox.sh.p11 or control AAV2-lox.sh.luc,
viral spread at the site of the injection was determined by RFP staining, an adjacent
slice was stained for Cre in brown, and p11 in blue. Right panel is a magnification of the
double staining in the infected area (white square), white arrows indicate cells
expressing Cre and p11 in control injected mice, yellow arrows indicate cells expressing
p11 alone and blue arrows indicate cells expressing Cre alone. Scale bar is 50 µm.
(d) Ten week old D1-Cre or D2-Cre mice were injected in the NAc with Cre inducible
AAV2-lox.sh.p11 or control AAV2-lox.shluc, wt littermates were injected with AAV2-sh.p11
or AAV2-sh.luc 10 weeks later mice were submitted to cocaine (7.5mg/kg) place conditioning
as described in the materials and methods section. Data (means ±SEM n=5–10
per group) were analyzed using two way ANOVA. Effect of injected virus F(1,
45)= 5.06, p=0.0301; effect of genotype F(2, 45)= 1.45,
NS; interaction F(2, 45)= 0.42, NS. Post hoc comparison (Tukey
HSD test) *p<0.05; **p<0.01.