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We used 4-HNE as a marker for senescent cells because it is closely associated with other markers of senescence in mouse liver including γH2A.X (Fig. 2F) and Sen-β-Gal (Wang et al., 2009, 2010) and allows clear cell boundary definition in the low magnification tiled images necessary for cluster analysis.
In a landmark study published in 1995, postdoc Goberdhan Dimri, Campisi, and other collaborators discovered a marker for senescent cells that allowed them to develop the first test to detect such cells in living tissue; they subsequently reported that senescent cells accumulate with age in humans.
However, the specificity and sensitivity of sen-β-Gal as a marker for senescent cells in vivo has been repeatedly questioned [ 47, 48].
There is currently no one single marker for senescent cells, but researchers now have characterized a number of important characteristics which have been summarized by Campisi and d'Adda di Faggana [ 9] and Cichowski and Hahn [ 10].
WFDC1 mRNA level was increased as well, implying yet again on its production by cells with reduced proliferation rate, and suggesting that WFDC1 level might be considered as a potential marker for senescent cells.
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Although there are no completely specific markers for senescent cells, many cell types express a pH-labile enzyme called senescence-associated β-galactosidase (SAβG) when they undergo senescence because of multiple stimuli.
Using sensitive and specific markers for senescent cells [ 51, 52], we found that short-term, adult-onset DR significantly reduced the frequencies of senescent liver hepatocytes, especially in the centrilobular area, and of senescent intestinal enterocytes in the transient amplifying zone.
Several recent studies have provided evidence that lamin B1 is a marker for senescence, and its expression is downregulated in senescent cells (Shimi et al., 2011; Freund et al., 2012; Dreesen et al., 2013).
They engineered mice so that when cells flipped on a gene called p16Ink4a, a marker for senescence, the cell would also turn on the production of inactive cellular death genes, not normally produced by senescent cells.
We did not observe calcification even though after 7 days of dox-treatment many other markers characteristic for senescent cells were present.
Senescence-associated β-galactosidase (SA-β-Gal) is a widely used marker of senescent cells in vitro and in vivo.
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