Supplementary Figure Legends
Supplementary Figure 1: A. RT-PCR analysis of PKC mRNA levels. Total RNA was
isolated from MCF10A, MCF10A-H1047R and MCF10A-E545K cell lines using
RNEasy Plus Mini Kits (Qiagen Inc., Mississauga, ON, Canada). cDNA from each cell
type was synthesized from 1.0ug total RNA using Qiagen Quantitect RT Kits. qPCR was
performed to amplify PKC cDNA and PUM1 cDNA (used as a reference) using the
Rotor-Gene SYBR Green PCR Kit and a Roche LightCycler thermocycler. PKC primers
were: 5’ GTC CGG GTG AAA GCC TAC TAC 3’ and 5’ ACG GGT CTC CTT CCT
CAT CT 3’. PUM1 primers were: 5’TGA GGT GTG CAC CAT GAA C 3’ and 5’ CAG
AAT GTG CTT GCC ATA GGG 3’. Standard curves were created for both PKC and
PUM1using RNA from MCF10A cells and used to determine relative amounts of PKC
and PUM1 mRNAs. Data shown are means  SE from three independent experiments.
B. MCF7 and MDA-MB-231 cells were either mock-transfected, transfected with a
control RNA duplex, or transfected with two different RNA duplexes targeting PKC. Six
days after transfection, cells were stained for SA-Gal activity. Percentages of SA-Gal
positive cells were determined as described in Materials and Methods. No basal or
induced SA-Gal activity was detected in MDA-MB-231 cells. Shown is a representative
example from three independent experiments.