Skip to main content
. Author manuscript; available in PMC: 2022 Jan 1.
Published in final edited form as: J Proteome Res. 2020 Nov 13;20(1):715–726. doi: 10.1021/acs.jproteome.0c00616

Figure 5: Validation of J.Lck.TurboID signaling in dialyzed serum.

Figure 5:

(A) J.Lck.TurboID cells were incubated in 0, 10, 30, 60, 120 minutes in 1 mM biotin in biotin-free RPMI containing different ratios of dialyzed:regular fetal serum with or without TCR stimulation (5 minutes) or pervanadate (PV) treatment as a positive control. ERK 1/2 pThr202/pTyr204 abundance was assessed via immunoblot, using the unphosphorylated ERK 1/2 as loading control. (B) J.Lck.TurboID, J.Lck.WT and JE6 cells were incubated in 30 minutes in 1 mM biotin in biotin-free RPMI media containing 2.5% regular serum and 7.5% dialyzed serum with or without TCR stimulation (5 minutes). ERK 1/2 pThr202/pTyr204 abundance was assessed via immunoblot, using the unphosphorylated ERK 1/2 as loading control. Biotinylation was assessed by streptavidin via immunoblot.