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. 2022 Aug 29;9:965921. doi: 10.3389/fmolb.2022.965921

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Copyright © 2022 Singh, Paschke, Tweedy and Insall.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Actin localizes to Ax2 and pkbA- protrusions, but not in pkgB- and pkbA-/pkgB-. (A) Subcellular localization of F-actin (Red). Life-act-mRFPmars2 was expressed in Ax2, and mutant cells. Cells were allowed to migrate under agarose mediated by a folate gradient and imaged by AiryScan confocal microscopy at a frame interval of 3 s (1f/3 s). Ax2 and pkbA- cells show F-actin in pseudopodia (asterisk). F-actin accumulation is absent in pkgB- protrusions, and highly reduced in pkbA-/pkgB-. Static F-actin in the cell body is present in both pkbA- and pkgB-. (B) Quantification of the florescence intensities in cells. Using ImageJ, intensity profile plots were generated by drawing a straight line across the cell. The amount of F-actin is similar in Ax2 and pkbA-, but reduced in pkgB- and pkbA-/pkgB- cells, where F-actin is mostly localized in the cell body. Images are representative of ≥10 cells from 3 independent experiments.