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. 2016 Oct 17;101(3):683–692. doi: 10.1189/jlb.2A0316-141RR

Figure 8. CXCL12-induced membrane ruffles and macropinocytic cups activate Akt.

Figure 8.

(A) BMMs expressing YFP-Akt-PH and CFP were imaged during CXCL12-stimulated macropinocytosis, and ratio images (YFP/CFP) were processed. The numbers at the top right in each frame indicate time following addition of CXCL12 (seconds). Red and white arrowheads indicate macropinocytic cups and corresponding YFP-Akt-PH signal, respectively. YFP-Akt-PH localized at lamellipodia and membrane ruffles (t = +0 to t = +80) then accumulated in macropinocytic cups (t = +120 to t = +220 s). Color bar indicates relative value of ratio intensities. Original scale bars, 3 μm. (B) Immunofluorescent staining of pAkt (Thr308) and Akt demonstrates that Akt was phosphorylated at the cup structure (arrowheads indicate macropinocytic cup) in a cell fixed 1 min after addition of CXCL12. Comparison of the phase-contrast image (phase) and the ratio image (pAkt/Akt) displays a strong ratio value at the macropinocytic cup. Color bar indicates relative value of ratio intensities. Original scale bar, 5 μm.