Figure 2.
Stable amber suppression in hiPSCs. A) Scheme for generating CTL07‐II‐AS, a hiPSC line with expanded genetic code, CpK incorporation in the GFP150TAG reporter and subsequent SPIEDAC fluorescent labeling with SiR‐Tetrazine. B) Brightfield and green fluorescence images of CTL07‐II and CTL07‐II‐AS. CTL07‐II‐AS cells that were incubated with 0.2 mM CpK for 24 h before imaging express GFP, while cells cultured without CpK and the parental cells do not show any fluorescence. C) SDS‐PAGE and fluorescent imaging of cell lysates of CTL07‐II and CTL07‐II‐AS, cultured in the presence or absence of 0.2 mM CpK for 24 h, incubated with 1 μM SiR‐Tetrazine to perform SPIEDAC labeling of CpK‐bearing proteins. Western blot shows FLAG‐PylRS expression in CTL07‐II‐AS cells. GAPDH was used as loading control.