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Supplemental Figure Legends
Supplemental Figure S1: Susceptibility of MDA-MB-231 cell line to CTL-mediated lysis
Cytotoxicity was determined by a conventional 4 h
51
Cr release assay at different effector: target
(E:T) ratios on 1000 target cells per well. Bars indicate the mean, and the error bars indicate the
standard deviations. Significant P-values are indicated by asterisks (* P=0.03; ** P=0.003; ***
P<0.001). (B) HLA-A2 molecule expression on the surfaces of MCF-7 and MDA-MB-231 cell lines.
Immunofluorescence staining was performed using MA2.1 monoclonal antibody and analysed on BD
LSR Flow Cytometer. (C) Lower expression of TAP1/2 in MDA-MB-231 cell line compared with
MDA-MB-231, as determined by western blot analysis. Total cell lysates (30 μg) were subjected to
SDS-PAGE, blotted, and probed with specific antibodies, as indicated. Equal protein loading was
assessed using β-actin.
Supplemental Figure S2: Side population (SP) analysis of epithelial and EMTed cell lines using the
DNA-binding dye (Hoechst 33342). The SP cells are indicated within the P2 gate.
Supplemental Figure S3: RT-qPCR showing relative mRNA expression of HLA-A2, TAP1, and
TAP2. Values were normalized to 18s rRNA levels. Error bars represent standard deviations.
Supplemental Figure S4: (A) Phosphorylated tyrosine (pTyr) accumulation at the immune synapse
formed between T cells and shWISP2 cells (blue: nucleus; green: pTyr). (B) Conjugates formed
between T cells and tumor cells were scored by visual counting using a confocal microscope. Error
bars represent standard deviations. Mean number of active synapses per field counted in 10 different
fields per cell line were randomly selected.
Supplemental Figure S5: Western blot analysis of the effect of Smad2 silencing on KLF-4
expression. Total cell lysates (25 μg) were subjected to SDS-PAGE, blotted, and probed with specific
antibodies, as indicated. Equal protein loading was assessed using β-actin.