Upon completion of the reaction, the substrate was rinsed with deionized water and dried at 60°C overnight and then heated at 420°C for 120 min.
Upon completion of the incubation period, substrates were washed gently with PBS and used for immunostaining.
Consistent with the published cell-based data, all four mutants exhibited elevated kinase activity compared to wild-type kinase as evident by the rapid completion of substrate monophosphorylation and appearance of diphosphorylated substrate peaks in the mass spectra of the oncogenic variants.
By contrast, when we used wild-type XBP1-BSL as the substrate, addition of Trl1 led to completion of the splicing reaction, consistent with the substrate's capacity to form ES1 and eject the intron (Fig 4E, lane 10).
This assay is conducted for an extended time period utilizing a high LSD1 concentration, with conditions where LSD1-catalyzed demethylation of the H3-21-K4Me2 substrate nears completion, resulting in extensive conversion of the substrate to mono- and unmethylated H3-21.
These indicator paints are observed upon completion of the deposition process to record the substrate temperature.
After completion of the direct imprint, the Al substrates are ultrasonically cleaned in acetone and alcohol solution.
Figure 6a,b,c,d presents the surface morphologies of the substrate after the completion of scratching with probe radiuses of 6, 8, 10, and 12 nm, respectively.
Figure 3 presents instantaneous defect structures and surface morphologies of the substrate after the completion of scratching and retraction for the two scratching depths.
Sil1-deficiency is predicted to cause accumulation of the ADP-bound form of BiP, which inefficiently releases substrate thus preventing completion of the folding process.
