Figure 5
Application of β1-tubulin–Venus reporter imMKCLs to siRNA-based assays. (A) Relative Venus intensity of β1-tubulin–Venus imMKCLs that were transfected with siRNAs specific for the indicated genes or nontarget. Cells were reversed transfected using the Stemfect RNA Transfection Kit and further matured for 7 days. Venus fluorescence intensities were detected using the ArrayScan and normalized to values of cells treated with mock siRNA in the presence of SR1 or DMSO. (B) Relative Venus intensity of β1-tubulin–Venus imMKCLs that were transfected with siRNAs specific for AhR, CYP1B1, or nontarget (Mock). Samples were also assessed for mRNA expression of AhR (C), CYP1B1 (D), or TUBB1 (E), as well as PLP yield (F). The mRNA expression levels were measured using quantitative real-time reverse transcription polymerase chain reaction and normalized to GAPDH. For PLP production, the cells were incubated for 7 days. CD41+CD42b+ PLPs were detected and counted using flow cytometry. Data are expressed as the mean ± SD from 3 independent experiments. *P < .05, **P < .01 vs Mock (without SR1), 1-way ANOVA. n.s., not significant (P > .5).

Application of β1-tubulin–Venus reporter imMKCLs to siRNA-based assays. (A) Relative Venus intensity of β1-tubulin–Venus imMKCLs that were transfected with siRNAs specific for the indicated genes or nontarget. Cells were reversed transfected using the Stemfect RNA Transfection Kit and further matured for 7 days. Venus fluorescence intensities were detected using the ArrayScan and normalized to values of cells treated with mock siRNA in the presence of SR1 or DMSO. (B) Relative Venus intensity of β1-tubulin–Venus imMKCLs that were transfected with siRNAs specific for AhR, CYP1B1, or nontarget (Mock). Samples were also assessed for mRNA expression of AhR (C), CYP1B1 (D), or TUBB1 (E), as well as PLP yield (F). The mRNA expression levels were measured using quantitative real-time reverse transcription polymerase chain reaction and normalized to GAPDH. For PLP production, the cells were incubated for 7 days. CD41+CD42b+ PLPs were detected and counted using flow cytometry. Data are expressed as the mean ± SD from 3 independent experiments. *P < .05, **P < .01 vs Mock (without SR1), 1-way ANOVA. n.s., not significant (P > .5).

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