Table 1.

PCR primers for the NPM and Bax mutants^a

Primer name	Sequence (5′–3′)
NPM (BamHI-EcoRI)
5′ Flag WT Fw	GAGCTCGGATCCGCCACCATGGACTACAAGGATGACGAGAAGATTCGATGGACATGGA
3′ Rev WT	GGATATCTGCAGAATTCTTAAAGAGACTTCCT
ΔNLS Fw	GCGCCACCAGTCTTAAGCTTGCGAGATACTCCAGCCAAAAAT
ΔNLS Rev	ATCTCGCAAGCTTAAGACTGGTGGCGCTTTTTCTTCAGCTTC
ΔNES Fw	GAAAAAGCTTTTAGCGTTGAAGTGTGGTTCAGGGCC
ΔNES Rev	CGCTAAAAGCTTTTTCTGTGGTATTTCAAAGCCCCCAAGG
K263R Mut	TGCCAGAGATCTTGAATAGCCTCTTGGTCAGTCATCCGGAAGCAATTCTTCACATAATTGATGAATTTGGCTTCCACTCTGGGAAGAGAACCAC
Bax (XbaI-BamHI-EcoRI)
V5 tag Fw	GAAGATTCTAGAGCCACCATGGGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACGAAGCTTATGGACGGGTCCGGGGAGCA
163A Fw	CGCGCCGGATCCAAGACCAGGGTGGTTGGGACGGCCTCCTCGCCTACTTTGGGACGCCCA
163E Fw	CGCGCCGGATCCAAGACCAGGGTGGTTGGGACGGCCTCCTCGAATACTTTGGGACGCCCA
184A Rev	GAGCTCGAATTCAGCCCATCTTCTTCCAGATGGTGAGTTCGGCGGTGAGCACTCCCGCCA
184E Rev	GAGCTCGAATTCAGCCCATCTTCTTCCAGATGGTGAGGGCGGCGGTGAGCACTCCCGCCA
WT Rev	GAGCTCGAATTCAGCCCATCTTCTTCCAGATGGT

^aShown is a list of forward (Fw) and reverse (Rev) primers used for generating wild-type (WT) NPM and Bax mutants using standard PCR (see Materials and Methods), including cytosol-restricted NPM mutant (NPM-ΔNLS) and nucleus-restricted NPM mutants NPM-ΔNES and NPM-K²⁶³R. Bax Ser¹⁶³ is the GSK3β-regulated site. Bax Ser¹⁸⁴ is the Akt-regulated site. V5 is an epitope tag. A and E refer to point mutations that render Bax either constitutively active or inactive, respectively, at specific serine sites.