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Similarly, PNP knockout experiments demonstrated the presence of fully processed collagen I and collagen II in tissues of experimental mice.
Collagenase digestion was used to confirm the intracellular location of fully processed collagen in the N extracts.
In contrast, SMase treatment decreased the amount of type II procollagen as well as resulting in a reduction in the level of processed collagen in the media.
However, the discovery of efficient intracellular procollagen processing in the absence of intracellular fibrils in postnatal tendon illustrates the ability of cells to prevent the premature assembly of processed collagen monomers into collagen fibrils [ 16].
The results show that BFA significantly reduced pNcollagen and fully processed collagen in the intracellular extracts of E13 chick metatarsal tendon, but pCcollagen was unchanged, consistent with the results obtained using the pulse chase protocol.
In E21 rat tail tendon, some intracellular collagen persisted following BFA treatment (see lanes marked 'BFA, N' in Figure 7A), even though the proportion of fully processed collagen was significantly decreased.
Similar(51)
Here we evaluated human adipose-derived stem cells (ADSCs) as a source of cells for heart valve tissue engineering investigating their ability to synthesize and process collagen and elastin.
Specifically, we recently demonstrated the diminution of osteoblastic differentiation on such a modified ECM [12] by using the lathyrogen β-aminopropionitrile (bAPN), an inhibitor of the enzyme lysyloxidase (Lox), which extracellularly processes collagen and initiates collagen cross-link formation [13].
MMP1 degrades collagen types I, II and III and the basement membrane proteins, MMP12 targets elastin, MMP8 and MMP13 can process collagen types I, II and III, while MT1-MMP can cleave a number of ECM proteins including fibronectin, laminin-1 and fibrillar collagen type I [ 70- 73].
This suggests a mechanism by which MMP-26 expression in early-invasive areas leads to activation of MMP-9, upon which then is capable to process collagen-4 and VEGF and thereby drives invasion and angiogenesis.
For collagen I, which is synthesized by fibroblasts as pro-collagen and processed to mature collagen (tropo-collagen) by cleavage of N- and C-terminal pro-peptides, COL1A1 mRNA and secretion of the C-terminal pro-peptide CICP were chosen as readouts.
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Justyna Jupowicz-Kozak
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