Temporal and spatial expression of alternative splice-forms of the α1(XI) collagen gene in fetal rat cartilage

Mutations within a furin consensus sequence block proteolytic release of ectodysplasin-A and cause X-linked hypohidrotic ectodermal dysplasia

“…Equal aliquots of cell extract or medium fraction were separated on 10% SDS͞PAGE gel and immunoblotted as described (31) with the use of chemiluminescence (ECL, Pierce).…”

Collagen 11a1 is indirectly activated by lymphocyte enhancer-binding factor 1 (Lef1) and negatively regulates osteoblast maturation

“…The Nterminal fragments V0 and V1B represent variations of the domain of Col11a1 that remains associated with the surface of collagen fibril for an extended period of time. V0 and V1B are the two most prevalent isoforms implicated in mineralization of the bone collar in long bone development (Davies et al 1998. In addition, the N-terminal fragments are proteolytically released from the surface of collagen fibrils slowly but at isoform-specific rates of processing (Thom et al 1991, Medeck et al 2003.…”

“…Protein aliquots were mixed with non-reducing protein sample dye and resolved by SDS 7% PAGE. Proteins were transferred to PVDF membranes for immunoblotting with a rabbit polyclonal antibody recognizing the C-telopeptide domain of collagen 11a1 (20545) (Li et al, 1995;Davies et al, 1998) or mouse monoclonal anti-decorin antibody (Scott et al, 1993). Col11a1 and decorin antibodies were diluted 1:1000 and 1:40, respectively, in TBST (TBS, 0.4% tween-20) with 3% BSA.…”

Expression, purification, and refolding of recombinant collagen α1(XI) amino terminal domain splice variants

“…Isoforms of the α1(XI) chain have been shown, by immunohistological and immunofluorescence detection, to distribute in a temporal, spatial and tissue specific manner with α1(XI) NTD[p7] and α1(XI) NTD[p6b+7] found primarily in cartilage [11] and [7]. During development of long bones and cartilage in rats, α1(XI) NTD[p7] is uniformly expressed while α1(XI) NTD[p6b+7] is restricted to the lateral boundaries of the diaphyseal cartilage, radius and ulna [12].…”

“…The most abundant forms of the α1 chain of type XI collagen are α1(XI) NTD[p7] and α1(XI) NTD[p6b+7] comprising 40% and 30% of expressed α1(XI) respectively (Fig. 1B) [11]. Isoforms of the pro-α1(XI) gene exhibit tissue-dependent temporal and spatial expression in rat fetal cartilage [12] and [13].…”