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. Author manuscript; available in PMC: 2023 Apr 6.
Published in final edited form as: Nat Cell Biol. 2022 Oct 6;24(10):1499–1515. doi: 10.1038/s41556-022-00997-7

Extended Data Figure 8: Global recruitment of Opto-ACTU+ can cause spatiotemporally confined activation of Ras/PI3K/Akt/TORC2/F-actin network components.

Extended Data Figure 8:

(A) Intensity profiles of LimE-GFP and Opto-ACTU+ along the white lines (the images are same as shown in Figure 6B) demonstrate that F-actin polymerizes in the domains of membrane where Opto-ACTU+ accumulates and when that leads to a protrusion, Opto-ACTU+ moves away with a short time delay. (B) 360° membrane kymograph of cell shown in Figure 6B. (C) 360° membrane kymograph of cell shown in Figure 6C. (D) Time-lapse live cell images of Dictyostelium cells co-expressing Opto-ACTU+, cAR1-CIBN, and PHCrac-YFP where recruitment was started at t=0s. Numbers show time in seconds. The “i” and “ii” are showing two different PIP3 production events which eventually lead protrusion formation. For each event, blue arrowheads are showing the areas where Opto-ACTU+ was first accumulated which in turn became the areas of PIP3 production and eventually, after protrusion formation, Opto-ACTU+ moved away to a newer back-state area. (E) 360° membrane kymograph of cell shown in (D). (F) 360° membrane kymograph of cell shown in Figure 6D.