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. 2023 Nov 9;24(22):16138. doi: 10.3390/ijms242216138

Figure 2.

Figure 2

Induction of YARS1 expression using the pSwitch-multi system in mammalian and invertebrate cell lines. Western blot analysis of protein extracts from (A) N2a, (B) CHO-K1, (C) HeLa, (D) HEK293T and (E) S2 stable and non-stable cell lines that have been transiently transfected with 1 µg of pUAST-attB-multi-YARS1. Four hours after transfection, a recommended concentration of Mifepristone (10 nM) was added to the medium in the conditions containing the two plasmids, for 3 days. (F) Western blot analysis of protein extract from S2 cells 24 h after transient co-transfection with pUAST-attB-multi-YARS1 and/or Actin-GAL4 driver (1:1 transfection ratio). Expression of exogenous YARS1 was detected using mouse monoclonal HA-tag antibody. Endogenous YARS1 expression could be detected in mammalian cells by using mouse monoclonal YARS1 antibody. Equal loading was validated by using mouse monoclonal α-tubulin antibody (n = 3). Note the exogenous YARS1 signal detection in the condition containing both vectors but without mifepristone induction in HeLa, CHO-K1 and HEK293T cells.