C6 software version 1

The FITC Annexin V Apoptosis Detection Kit I (BD Bioscience) was used. Cell lines treated with the reagents for 4 days were suspended in binding buffer and incubated with FITC-labeled annexin V and propidium iodide in the dark.
Flow cytometric measurements were performed on a BD Accuri C6 Flow Cytometer (BD Biosciences). A 488-nm blue laser was used for excitation, and signals were detected using the FL1 channel (533 nm) for FITC and the FL2 channel (585 nm) for propidium iodide. The signals of 30,000 events were obtained. Analyses of the obtained data were performed using C6 software version 1.0 (BD Biosciences).

The FITC Annexin V Apoptosis Detection Kit I (BD Biosciences, San Jose, CA, United States) was used. Cell lines treated with doxycycline for 4 days were suspended in binding buffer and incubated with FITC-labeled annexin V and propidium iodide in the dark.
Flow cytometric measurements were performed on a BD Accuri C6 Flow Cytometer (BD Biosciences, San Jose, CA, United States). A 488-nm blue laser was used for excitation, and signals were detected using the FL1 channel (533 nm) for FITC and the FL2 channel (585 nm) for propidium iodide. The signals of 30,000 events were obtained. Analyses of the obtained data were performed by using C6 software version 1.0 (BD Biosciences, San Jose, CA, United States).

Pyrimidine salvage activity was estimated based on the availability of EU. Cells were incubated in the medium with EU for 3 h after treatment with teriflunomide for 72 h. EU incorporated in RNA was labeled with Alexa Fluor 488 using the Click-It RNA Alexa Fluor 488 Imaging Kit (Thermo Fisher Scientific, Waltham, MA, USA). The signal for EU was measured using flow cytometric measurements performed on a BD Accuri C6 Flow Cytometer (BD Bioscience, Franklin Lakes, NJ, USA). A 488-nm blue laser was used for excitation, and the signal was detected by the FL1 channel (533 nm). The signals of 30,000 events were obtained. Analyses of the obtained data were performed using C6 software version 1.0 (BD Biosciences).