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In contrast pLB cell proliferation is suppressed by neighboring pLUM or MCF7 cells, which also involves secreted factors.
One of these areas was close to the cell body of the PeD12 cell, and the other was close to the PlB cell body.
The PLB cell was mounted on a separate bracket attached to a micrometer-controlled translation stage to allow focusing, and the stage was again bolted to the optical rail.
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Lee et al. (2013) demonstrated that during PLB formation, there is a transient formation of storage proteins within PLB cells.
Unless otherwise specified, for lysing granulocytes and PLB cells, 4x Laemmli sample buffer was mixed with equal volume of distilled water (dH2O) supplemented with 5 mM EDTA (250 mM stock in dH2O, pH 7.4 with NaOH) and protease inhibitor cocktail (1 Complete Mini tablet in 10 ml dH2O, Roche Applied Science, www.roche-applied-science.com).com
As mixed cells segregate by subtype within colonies and tumors with minimal direct pLUM and pLB cell-cell contact, we speculated that pLUM suppress pLB via a diffusible factor.
Volcano plot of differentially expressed genes in pure luminal (pLUM) versus pure luminobasal (pLB) cells.
We show that pLUM and MCF7 cells suppress proliferation of pLB cells in mixed-cell 3D colonies in vitro and that pLUM cells suppress growth of pLB cells in mixed-cell xenografts in vivo.
Since pLUM cells suppress pLB cells the 1 1 ratio was chosen so that sufficient numbers of pLB cells would remain in 6-week-old tumors to allow their quantitation despite further EGFRi suppression.
pLB cells are propagated in vitro under EWD conditions; pLUM cells are propagated under E-replete conditions.
For that we needed pure luminal-derived ER PR CK5+ cells for drug screening, that is, the pLB cells reported here.
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