Immunoblotting and Chromatin Immunoprecipitation Assays

This was performed as described previously.^{35 (link), 36 (link)} Antibodies used in WB included mouse monoclonal antibodies against β-actin (Sigma), α-tubulin (Sigma), E-cadherin (610404, BD), p-HSF-1/S326 (ab76076, Epitomics/Abcam), p-JNK/S63/73 (sc-6254; Santa Cruz), as well as, rabbit antibodies against Slug (AP2053a,ABGENT), HSF-1 (4356, Cell Signaling), Hsp70 (4876, Cell Signaling), p38 (9212, Cell Signaling), p-p38/T180/Y182 (4511, Cell Signaling), pERK/T202/Y204 (9109, Cell Signaling), Akt/pan (4691, Cell Signaling), p-Akt/S473 (4060, Cell Signaling), mTOR (2983, Cell Signaling), p-mTOR/S2448 (5536, Cell Signaling), JNK (sc-571, Santa Cruz), HER2 (2165, Cell Signaling), p-HER2/Y1278 (2247, Cell Signaling). Rabbit polyclonal HSF-1 antibody used in IP and ChIP assay was from Cell signaling (4356).

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Carpenter R.L., Paw I., Dewhirst M.W, & Lo H.W. (2014). Akt phosphorylates and activates HSF-1 independent of heat shock, leading to Slug overexpression and epithelial-mesenchymal transition (EMT) of HER2-overexpressing breast cancer cells. Oncogene, 34(5), 546-557.

Publication 2014

β-actin α-tubulin E-cadherin p-HSF-1/S326 p-JNK/S63/73 AP2053a Hsp70 p-p38/T180/Y182 pERK/T202/Y204 Akt/pan p-Akt/S473 p-mTOR/S2448 JNK (sc-571, p-HER2/Y1278

Actin Antibodies Antibody Chip assay E cadherin Her2 Hsp70 Monoclonal antibodies Mouse Mtor Rabbit Slug α tubulin

Corresponding Organization :

Other organizations : Duke University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of β-actin
Protein expression levels of α-tubulin
Protein expression levels of E-cadherin
Phosphorylation levels of HSF-1/S326
Phosphorylation levels of JNK/S63/73
Protein expression levels of Slug
Protein expression levels of HSF-1
Protein expression levels of Hsp70
Protein expression levels of p38
Phosphorylation levels of p38/T180/Y182
Phosphorylation levels of ERK/T202/Y204
Protein expression levels of Akt/pan
Phosphorylation levels of Akt/S473
Protein expression levels of mTOR
Phosphorylation levels of mTOR/S2448
Protein expression levels of JNK
Protein expression levels of HER2
Phosphorylation levels of HER2/Y1278

control variables

None explicitly mentioned

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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