Figure 3

Stimulation and inhibition of STAT3 phosphorylation detected with HiBiT-BRET approach using HiBiT/CRISPR-modified cells. (a) Pooled K562 STAT3-HiBiT cells (60,000 cells/well) were treated with different cytokines at 0.1 µg/mL final concentration in different time points. (b) 60,000 pooled K562 STAT3-HiBiT cells seeded overnight were treated the next day with 0.1 µg/mL IL-27 every 10 min for 90 min. (c) K562 STAT3-HiBiT Cells (Clone) (60,000 cells/well) or (d) LNCaP STAT3-HiBiT pooled population cells (40,000 cells/well) were treated with serially diluted IL-27 or IL-6, respectively for 40 min at 37 °C. (e,f) Dose-dependent inhibition of STAT3 phosphorylation with ruxolitinib in K562 or LNCaP STAT3-HiBiT/CRISPR-modified cells. Results are representative of at least two independent experiments performed in duplicates.