Test digestion protocol
The expected DNA insert sizes from pHACE-PKC plasmids digested by XhoI were shown.
This XhoI fragment contains a fully functional cDNA including a start codon, a PKC ORF and a stop codon.
  
PKC Constructs
PKC-HA
.
Vectors
pHACE or pHACB
 .
PKC Mutants
WT
DN
DPS
DNPS
CAT
CAT-KR
  

Mutation (Diagram)
wild type
kinase inactive
pseudosubstrate sequence deletion
N-terminal pseudosubstrate domain deletion
catalytic domain
catalytic domain mutant (kinase inactive)
  
PKCa
2.1kb
 2.1kb   2.0kb 1.1kb
1.1kb
  
PKCb1
2.1kb
 2.1kb   2.0kb
1.1kb
-
  
PKCb2
2.1kb
 2.1kb   2.0kb
1.1kb
-
  
PKCg
 2.2kb 2.2kb   2.1kb
1.1kb
-
  
PKCd
 2.1kb 2.1kb   1.5kb
1.1kb
1.1kb
  
PKCe
 2.3kb 2.3kb   1.8kb
1.1kb
1.1kb
  
PKCh
 2.1kb 2.1kb   1.6kb
1.1kb
-
  
PKCz
 1.8kb 1.8kb   1.5kb
1.1kb
1.1kb
  
PKCi
 1.8kb 1.8kb   1.5kb
1.1kb
-
  
Data
 Structures of the PKC mutants Structure of pHACE-PKC constructs  
Expression level in COS-7 cells Coding sequences of the PKC mutants
Western blot data Kinase assay data
Immunofluorescence microscopy Test digestion protocol

WT: wild type

KR: kinase inactive mutant (dominant negative)

DPS: pseudosubstrate sequence deletion mutant (constitutively active)

DNPS: N-terminal pseudosubstrate domain deletion mutant (constitutively active)

CAT: catalytic domain mutant (constitutively active)

CAT-KR: catalytic domain mutant (kinase inactive)