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Both methods are comprehensively compared using assorted matrices derived from engineering applications.
Cells were cultured overnight in the assorted matrices and lysates were prepared as previously described [ 45, 51].
Motility experiments using the assorted matrices were performed a minimum of 3 times using different batches of matrices and following a minimum of 10 individual cells.
The role of FAP in the matrix-induced permissive tumor behavior was assessed in Panc-1 cells in assorted matrices by time-lapse acquisition assays.
When activity levels were compared for Panc-1 cells cultured within the assorted matrices in the presence or absence of mAb13, a significant down-regulation of constitutive FAK activity levels under β1-integrins inhibition was observed in all three matrices (p = 0.02, p = 0.002, p = 0.02 for FAP-, FAP+inhibitoribitor matrices, respectively).
Similar(55)
Briefly, 12 well plates containing assorted 3D matrices were used and 8,000 MDA-MB-231 cells were cultured overnight in the presence or the absence of inhibitors or respective negative controls (for details see Inhibitors above).
To determine the downstream components of β1-integrins, Panc-1 cells were cultured within assorted 3D matrices and the constitutive activity levels of both AKT and FAK were analyzed by the ratios of pS-AKT/total AKT and pY-FAK/total FAK, respectively.
The levels of ECM proteins (tenascin C, collagen I and fibronectin) and cellular α-SMA potentially underlying the matrix differences mediated by FAP were measured on the assorted un-extracted matrices.
Based on this process of the mandibular development and growth, Moss and Simon assorted functional units using functional matrix theory [13].
After removing fibroblasts, extracted matrices were re-cultured with breast epithelial cells of assorted characteristics: MCF-10A (non-tumorigenic), MCF-7 (tumorigenic, non-invasive), and MDA-MB-231 (tumorigenic, invasive).
We observed that assorted breast epithelial cells reacted differently to two-dimensional vs. staged, control (early) and tumor-associated (late), three-dimensional matrices.
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