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Furthermore, investigating the effects of added growth factors, such as Gdnf and Gdf11, on isolated Fras1 mutant MDs stripped of their surrounding mesenchyme (45) may in future add extra mechanistic insights beyond those obtain from the renal tract cultures, as described in this study.
In cloned transfectants derived from both lines, PAX3-FKHR expression resulted in increased proliferative rate in vitro and promoted cell growth in the absence of added growth factors.
For the first 3d, Mixl1GFP/w cells (responder cells) were differentiated in the absence of added growth factors.
Mouse aortic rings were cultured in collagen gels with serum, but otherwise in the absence of added growth factors.
Importantly, α-SMA siRNA also significantly reduced α-SMA protein levels as measured by Western blot (Figure 10b); again, this reduction was effective even in the presence of added growth factor, and non-specific siRNA control elicited no such decrease in target protein.
Importantly, CCT-eta siRNA also significantly reduced CCT-eta protein levels as measured by Western blot (Figure 3c); again, this reduction was effective even in the presence of added growth factors, and control scrambled siRNA elicited no such decrease in target protein.
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Next, we examined the effects of adding growth factors HGF, FGF1, FGF7, and FGF10, to the culture medium of cancer cells.
We next examined the effects of adding growth factors to the culture medium of cancer cells and the effects of FGFR inhibitor and MET inhibitor treatment on cell proliferation induced by fibroblast supernatant.
The results of experiments to evaluate the effects of adding growth factors and kinase inhibitors suggests that the stimulating effect of fibroblasts was attributable in part to HGF/MET or FGF/FGFR.
These results indicate that cells in this system are still capable of proliferating in vitro in the absence of exogenously added growth factors, suggesting the possibility of growth factor(s) being endogenously synthesized in these cultures.
These cells act as a source of endogenous signalling and preclude determination of direct or indirect action of exogenously added growth factors.
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