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. 2021 Apr 7;9:657456. doi: 10.3389/fcell.2021.657456

FIGURE 1.

FIGURE 1

Proteins from ANL-treated MetRS*MSCs can be detected via alkyne-Cy7 click-staining. MSCs were transfected with Lenti-CAG-MetRSL274G–mCherry (MetRS*MSCs) or a control lentiviral vector (CTLMSCs). (A) Morphology of the transfected cells was evaluated via phase contrast microscopy (top), and transfection of MetRS*MSCs was confirmed by visualizing mCherry fluorescence (bottom). (B,C) MetRS*MSCs and CTLMSCs were incubated with ANL (+) or vehicle (–) for 24 h. (B) MSCs were lysed; then, proteins were separated on an SDS-PAGE gel, click-stained with alkyne-Cy7, and imaged in stain-free and Cy7 mode (BONCAT). (C) MSCs were permeabilized, click-stained with alkyne-Alexa Fluor 488, and viewed via phase contrast microscopy (top) or under fluorescence (bottom) (FUNCAT); nuclei were counterstained with Hoechst, and ANL-labeled proteins were identified via alkyne-Alexa Fluor 488 fluorescence.