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Fig. 3 | BMC Biology

Fig. 3

From: Steering cell migration by alternating blebs and actin-rich protrusions

Fig. 3

Protrusion formation and orientation in ezrin morphant mesendoderm cells. a Exemplary ezrin-MO-injected mesendoderm cells displaying blebs (black arrowheads). Cells express Lifeact-GFP (green) and Dextran-Alexa 594 (red). Scale bar = 10 μm. b, c Quantification of bleb formation frequency (b) and bleb size at maximal expansion normalized to cell size (c) in control and ezrin-MO-injected mesendoderm cells. Note that bleb frequency also includes the false negatives not detected by APA (Additional file 4: Figure S2). d Orientation of actin-rich protrusion formation in ezrin-MO-injected cells with respect to the local direction of migration. The arrows below the diagrams indicate the direction of migration. The orientation of actin-rich protrusions was weighted by their actin content (i.e., total Lifeact fluorescence) to account for size differences between protrusions, their number is thus given in arbitrary units. POP: mean ± SEM of the magnitude of the polar order parameter. e Ratio of tumbling to run times in migrating single lateral ezrin morphant mesendoderm cells (ezrin-MO). Cells were tracked during the approximately first 2 hours after transplantation. The ratio was normalized to transplanted control cells in the same embryo (internal controls) to account for experimental variability between different embryos. Number of analyzed cells in (b, d) = 17 for control and 6 for ezrin-MO; (e) = 21 for ezrin-MO. Number of blebs in (c) = 19 for control and 21 for ezrin-MO. Statistical significance by Mann–Whitney test (b, c), by non-overlapping SEM of the POP (d) (see also Additional file 7: Figure S3D) or by one-sided t-test (e)

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