Fig. 7. Protein Kinase C activation in NA and TA cells. (A) NA and TA cells were
serum starved for 2 days then either left untreated or treated with 10 ng/ml
EGF or 100 ng/ml TPA for 5 minutes. Cells were scraped into lysis buffer and
soluble (S) and insoluble (I) fractions were prepared as described in the
Materials and Methods. Equal amounts of each fraction were electrophoresed on
10% SDS-PAGE gels and analysed by western blotting using antibodies
recognising PKC
, PKC
, PKC
and PKC
. (B) PKC
phosphorylation in NA and TA cells. Whole cell extracts were prepared from NA
and TA cells that had been serum starved for 2 days then left either untreated
or treated with 10 ng/ml EGF or 100 ng/ml TPA for 5 minutes. Western analysis
followed using polyclonal antibodies recognising phosphoPKC (pan) antibody,
which, in A431 cells, recognises phosphoPKC and phosphoPKC
only.
This was confirmed by probing adjacent lanes with PKC and PKC
antibodies and showing co-migrating bands (data not shown).
