Figure 1

Effect of pentazocine (P), haloperidol (H) and guanine nucleotides on the interaction between the Sig1R, Rac1, and Bcl-2. Mitochondria were incubated with sigma ligands and guanine nucleotides as described in Material and methods. Mitochondrial extracts were immunoprecipitated with anti-Sig1R (IP: Sig1R, A, B, E, Fa, Fb), with anti-IP3R (IP: IP3R, C,D) or anti-Bcl2 (IP: Bcl2, Fc) antibodies, the immunoprecipitate was subjected to SDS-PAGE and transferred. The blots were then probed to Western analysis with antibodies against Rac1(Wb: Rac1, A,C,Fc), Bcl2(Wb:Bcl2, Ea), MFN2(Wb:MFN2, Eb), IP3R (Wb:IP3R, Fa) and BiP (Wb:BiP, Fb). Results are representative of three independent experiments. (A, C, E, ) Lane 1 – control, no additions; lane 2 – plus GppNp; lane 3 – plus GDP; lane 4 – plus haloperidol; lane 5- plus (+)-pentazocine; lane 6- plus haloperidol and (+)-pentazocine. (F) Lane 1 – no additions; lane 2 – plus haloperidol, lane 3 – plus (+)-pentazocine; lane 4 - plus haloperidol and (+)-pentazocine. (B) Quantification of Rac1 blots shown in A; n = 3. *P < 0.05, compared with control levels. (D) Quantification of Rac1 blots shown in C; n = 3. *P < 0.05, compared with control levels.