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

Figure 3.

Figure 3

Concentration of inducible plasmid to be transfected in stable cell lines for pSwitch-multi regulatory vector. N2a cells stably expressing the pSwitch-multi regulatory vector have been transiently transfected with increased concentrations of inducible plasmid (ranging from 0 to 1 µg) containing (A) YARS1 or (B) AARS1 cDNA sequence. Four hours after transfection, a recommended Mifepristone concentration (10 nM) was added to the medium to induce transgene expression for 24 h. The transgene signal was first determined by using the HA-tag antibody. Exogenous and endogenous YARS1 and AARS1 expression was detected with mouse monoclonal YARS1 or AARS1 antibodies, respectively. Equal loading was validated by using mouse monoclonal α-tubulin antibody. Each graph represents the relative quantification of each band’s intensity compared to the expression level at 0.5 µg inducible plasmid without the presence of Mifepristone (n = 3). Bar charts are represented with s.e.m. for endogenous (black), exogenous in absence (grey) or presence of Mifepristone (red) YARS1 and AARS1, respectively. Statistical significance (** p < 0.01, **** p < 0.0001, ns—not significant) was determined using one-way ANOVA analysis.