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In agreement with these data, pyramidal neurons recorded in DAF-2DA-loaded slices from either mutant or control mice did not exhibit any fluorescence (Fig. 7).
Yet, when cell death is inhibited, DKO myotubes can differentiate as efficiently as Rb mutant or control cultures, demonstrating that neither factor is required for stimulating or maintaining the differentiation program.
Incubating brain slices for 30 minutes in a ringer solution containing 1400W (3 µM) had no effect on the punctate, neuronal fluorescence in slices from either mutant or control mice (Fig. 4, 30 sec).
The neuronal, punctuate DAF-2DA fluorescence in either mutant or control slices was not affected by the thapsigargin pre-incubation (Fig 5A), in agreement with data showing that neuronal NO production depends mainly on Ca2+ entering through NMDA channels [25], [26].
ESC-like cell were detected several days later and established as mutant or control rESCs.
There was little detectable BrdU incorporation at 6 dpf in either mutant or control populations.
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Cell lysate was prepared from subconfluent monolayer culture of ECs at 72 h post transduction of R-Ras mutants or control vector.
Many differentiation genes (Mef2c, Alx1, Alx4, and Alpl) exhibit significantly greater expression in Fgfr1 mutants compared to Pdgfra mutants or control embryos, consistent with the MEPM data in which these genes display greater expression at 4 hours following PDGF treatment compared to 4 hours following FGF treatment.
(B ) Immortalized MEFs from Lxrα − / − Lxrβ − / − mice reconstituted with wild-type human LXRα, AF1-deletion mutant (∆AF1), DNA-binding domain deletion mutant (∆DBD), ligand-binging domain deletion mutant (∆LBD), AF2-deletion mutant (∆AF2) or control mock were pretreated with the LXR agonist GW3965 (1 µM) overnight, followed by stimulation with LPS (10 ng/ml) for 4 hr.
Hybrid animals bearing Canton S and ve e (mutant or parental control) were used to control for second site recessive mutations.
Since purified protein assays showed that W32S substitution lowers the propensity of SOD1 to aggregate in the presence of competent SOD1 seed, we next explored this phenomenon in living cells using a co-transfection system where one vector encoded a fluorescent reporter of inclusion formation and the other induced inclusion formation (untagged mutant SOD1 or control).
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