10.6084/m9.figshare.c.3604745_D5.v1 Hwanhee Kim Hwanhee Kim Jun-Young Oh Jun-Young Oh Sun-Lim Choi Sun-Lim Choi Yeon-Ju Nam Yeon-Ju Nam Anna Jo Anna Jo Ara Kwon Ara Kwon Eun-Young Shin Eun-Young Shin Eung-Gook Kim Eung-Gook Kim Hyong Kim Hyong Kim Additional file 6: Figure S6. of Down-regulation of p21-activated serine/threonine kinase 1 is involved in loss of mesencephalic dopamine neurons Springer Nature 2016 PAK1 Bcl-2 Apoptosis Neurodegeneration Parkinson’s disease 2016-04-27 05:00:00 Figure https://springernature.figshare.com/articles/figure/Additional_file_6_Figure_S6_of_Down-regulation_of_p21-activated_serine_threonine_kinase_1_is_involved_in_loss_of_mesencephalic_dopamine_neurons/4343696 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)