Do I need to change S4U-containing medium every 3 hours?

When performing longer duration labeling experiments, supplying fresh S4U-containing media every 3 hours enhances the S4U incorporation into the cells. As incorporation rates differ among cell types it is best to assess this directly using the SLAMseq Explorer Kit – S4U Incorporation Module. Figure 1 depicts an example for mouse embryonic stem cells (mESC), where different frequencies of S4U-containing media exchange were compared to no media exchange (Figure 1).

Figure 1 | S4U Incorporation rates for mouse embryonic stem cells (mESC). Media exchange every 3, 6, or 8 hours, was compared. Cells without media exchange show reduced incorporation rates particularly after 12-14 hours while 3-hourly media exchange provides the highest S4U incorporation rate over time.