Figure 2 | Scientific Reports

Figure 2

From: Rare CASP6N73T variant associated with hippocampal volume exhibits decreased proteolytic activity, synaptic transmission defect, and neurodegeneration

Figure 2

Self-processing of recombinant Casp6N73T zymogen. (a) Schematic diagram of recombinant human Casp6 zymogen showing the subunits and epitopes recognized by antibodies. The epitope for the SC-81635 antibody is unknown but located in the large subunit (LS), neoepitope for 10630 antisera detects the C-terminus of the LS of Casp6, and epitope for the Pharmingen antibody is G271-K285. Pro pro-domain, LS large subunit, L linker, SS small subunit. (b) Coomassie blue stain of 2 µg purified recombinant Casp6WT and Casp6N73T zymogens processed into LSL, LS and SS subunits. The unprocessed Casp6D(23,179,193)A was used as a control for the FL form. LSL large subunit with the linker, FL full length. * indicates unspecific band. (c) Quantification of the density of LSL, LS or SS normalized to density levels of Casp6WT from Coomassie blue stained gels. (d,f,h) Western blot of Casp6WT, Casp6N73T, and Casp6D(23,179,193)A with SC-81635 antibody anti-LS (d), with neoepitope antisera 10630 anti cleaved LS (f), and with Pharmingen antibody anti-SS (h). Quantification of (e) LSL and LS from (d), (g) LS from (f), and (i) SS from (h). Data represents mean and s.e.m of 3 independent experiments. No statistical differences were found by unpaired student’s test.

Back to article page