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. 2020 Apr 8;12(4):909. doi: 10.3390/cancers12040909

Figure 4.

Figure 4

MAPK phosphatase 1 (MKP1) is responsible for SIRT1-dependent ERK dephosphorylation. (A) IB analysis of MKP1 expression and acetylation in 293T cells. Cells were transiently transfected with indicated vectors and then immunoprecipitation (IP) was performed with an anti-pan-acetyl lysine (acK) antibody, followed by IB with an anti-Myc antibody. IgG, a negative control for IP. (B) IB analysis of acetylation status of MKP1 upon overexpression of p300 and/or SIRT1 in 293T cells. IP was performed with an anti-Myc antibody, followed by IB with an anti-pan acK antibody. (C) IB analysis of phospho-ERK levels depending on SIRT1 expression in the presence or absence of a MKP1/6 inhibitor (E/Z)-BCl Hcl in Dox-exposed Ctrl-iRas (Lane# 1, 3, 5, and 7) and Sirt1-iRas (Lane# 2, 4, 6, and 8) cells. MKP1/6 inhibitor (E/Z)-BCl Hcl and Dox was treated for 3h. ERK2 and α-tubulin, protein loading controls for IB. The whole western blot images please find in Figure S7.