Title and legend to supplementary figures

Supplementary Figure 1 Downregulation of LDHA decreases glycolysis.

a, MCF7/ErbB2 cells were transfected with scramble siRNA (Control) or LDHA siRNA as described under “Materials and methods”. 48 hours after siRNA transfection, cell lysates were prepared and immunoblot analyses were carried out with antibodies against LDHA and β-Actin. b, 48 hours after siRNA transfection, cells were transferred to 24-well plates for glucose uptake assay. Data are shown in percentage relative to control-transfected cells. c, 48 hours after siRNA transfection, cells were collected for LDH activity assay. Data are shown in percentage relative to control-transfected cells. Columns, mean of three independent experiments; bars, SE. **, P<0.01, ***, P<0.001.

Supplementary Figure 2 Downregulation of LDHA inhibits cell growth.

a, BT474M1 cells were transfected with scramble siRNA (control) or LDHA siRNA. 48 hours after siRNA transfection, cell lysates were prepared and immunoblot analyses were carried out with antibodies against HSF1 and β-Actin (insert). b, 48 hours after siRNA transfection, cells were plated into a 24-well plate at 5 x 103 cells per well on day 0. Then cells were counted at day 2, 4 and 6.

Columns, mean of three independent experiments; bars, SE. ***, P<0.001.

Supplementary Figure 3 Herceptin inhibits ErbB2-mediated induction of HSF1.

BT474M1 cells were treated with 0, 25, 50, 100μg/ml Herceptin (HCP) for 24h. The cell lysates were prepared and Western blot analyses were carried out with antibodies against ErbB2, HSF1 and β-Actin (top). Results were quantified by software Scion Image and relative intensity of HSF1 was calculated using the protein level without HCP treatment as 1.00 (bottom). Columns, mean of three independent experiments; bars, SE.

Supplementary Figure 4 Heregulin induces LDH-A expression by up-regulating HSF1.

MCF7 cells were transfected with scramble siRNA (Control) or HSF1 siRNA. 24 hours after siRNA transfection, cells were serum starved for 24h and then treated with 25ng/μl heregulinβ1 (HRG β1) for 24h. Cells were then lysed and extracts probed for HSF1 and LDHA. β-Actin was probed as a control for equal loading.

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