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. 2019 Jul 24;294(37):13671–13680. doi: 10.1074/jbc.RA119.008548

Figure 3.

Figure 3.

AKT mediates phosphorylation of IGPR-1 at Ser-220. A, equal number of PAE cells expressing IGPR-1 were seeded on the cell culture plates for various time points. The cells were lysed, and whole-cell lysates were blotted for phospho-AKT, total AKT, and GAPDH. The graph is representative of three independent experiments. B, equal number of PAE cells expressing EV or IGPR-1 alone or co-expressed with mutant dominant-negative AKT1 (K179M/T308A/S473A) were lysed, and whole-cell lysates were blotted for pSer-220 (pSer220), total IGPR-1, and AKT. C, IGPR-1/PAE cells were treated with control vehicle or GSK-690693 for 60 min, the cells were lysed, and whole-cell lysates were blotted for pSer-220, total IGPR-1, pSer-9–GSK3β, total GSK3β, and GAPDH. The graph is representative of three independent experiments. pSer-220 level is normalized to total IGPR-1. D, HUVECs were treated with control vehicle or GSK-690693 for 60 min, and whole-cell lysates were blotted for pSer-220, total IGPR-1, or GAPDH. The graph is pSer-220 level normalized to total IGPR-1, which is representative of three independent experiments. Mut. or mut., mutant.