Cell Line Generation and Culture Protocol

MCF7, MDA-MB-231, SK-BR-3 and IMR90 cells were purchased from American Type Culture Collection (ATCC). MDA-MB-231-GRP78^ind cells [8 (link)] (Supplementary Figure 7), and human iPSC lines [26 (link), 27 (link)] were generated as previously reported. All cells were cultured as described previously [8 (link)]. DCD-FLAG construct (ORF clone OHu30172) was purchased from Genscript. M50 Super 8x TOPFlash and M51 Super 8x FOPFlash was a gift from Randall Moon (Addgene plasmid #12456). pHRL-SV40 was obtained from Promega. CHIR99021 was purchased from StemCell Technologies (#72054). XAV939 and ROCK inhibitor Y-27632 were purchased from Sigma (#X3004 and #SCM075). GRP78-N20 (#sc-1050) and DCD-N20 (#sc-27465) antibodies, or Goat-IgG controls (#sc-2028), used in inhibitory experiments, were purchased from Santa Cruz Biotechnology.
Remaining Materials and Methods are available as a Supplementary File.

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Lager T.W., Conner C., Keating C.R., Warshaw J.N, & Panopoulos A.D. (2021). Cell surface GRP78 and Dermcidin cooperate to regulate breast cancer cell migration through Wnt signaling. Oncogene, 40(23), 4050-4059.

Publication 2021

MCF7 MDA-MB-231 SK-BR-3 IMR90 DCD-FLAG construct (ORF clone OHu30172) pHRL-SV40 CHIR99021 XAV939 and ROCK inhibitor Y-27632 GRP78-N20 (#sc-1050) and DCD-N20 (#sc-27465) antibodies Goat-IgG controls (#sc-2028),

Antibodies Cells Chir99021 Clone Goat Grp78 Human Inhibitory Ipsc Mcf7 Mda mb 231 cells Plasmid Promega Stemcell Sv40 Xav939 Y 27632

Corresponding Organization :

Other organizations : University of Notre Dame

Top 5 similar protocols

Variable analysis

independent variables

DCD-FLAG construct
M50 Super 8x TOPFlash
M51 Super 8x FOPFlash
CHIR99021
XAV939
ROCK inhibitor Y-27632
GRP78-N20 antibody
DCD-N20 antibody
Goat-IgG controls

dependent variables

Cell responses and outcomes

control variables

MCF7, MDA-MB-231, SK-BR-3 and IMR90 cells
MDA-MB-231-GRP78^ind cells
Human iPSC lines

positive controls

M50 Super 8x TOPFlash
M51 Super 8x FOPFlash

negative controls

Goat-IgG controls

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