Additional file 6: Figure S6. of Down-regulation of p21-activated serine/threonine kinase 1 is involved in loss of mesencephalic dopamine neurons

Other mutants of PAK1- inactive form reduce Bcl-2 level. a. HEK cells were transfected with 1, 2, or 3 μg of GFP, PAK1-WT, PAK1-H83,86L, or PAK1-K299R vectors and incubated for 48 h. The cell lysates were subjected to Western blotting analysis using anti-Myc, Bcl-2, Bcl-xL, or GAPDH antibody. b. The autoinhibitory domain (AID) of hPAK1 (67–150 amino acids) were amplified by PCR methods (hPAk1-67-R1-S: 5′-ggaattcggaagaaagagaaagagcggc-3′, hPAK1-150-Xho-A: 5′-ccgctcgagttaagctgacttatctgtaaagc-3′) and inserted to EcoRI/XhoI site of pCMV-Myc vector (Clontech) in-frame with Myc-tag, producing PAK1-DN (67–150) [47]. HEK cells were transfected with 1, 2, or 3 μg of PAK1-DN (67–150) vector and cell lysates were analyzed by Western blotting assay. (TIF 278 kb)