Figure 6. Distribution of Arp2/3, WAVE, and WASP at the Leading Edge(A-C) Top: representative images of GFP-tagged ARX-2 (A), WVE-1 (B), and WSP-1 (C) in knockin animals. Left, GFP knockin fluorescence (green); middle,mCherry-tagged plasma membrane and histone (red, plot region is labeled with a dashed line); right, merged images. Bottom: the GFP fluorescence distributionplot along the leading edge of the representative image shown above. The GFP fluorescence intensity along the leading edge was normalized to 1.5 times that inthe adjacent
hyp7 cell. The area above the baseline of 1.0 is highlighted in green. Individual GFP::WSP-1 puncta are numbered in (C). Scale bar, 2.5 um.(D) The leading-edge distribution of ARX-2::GFP. Left, merged images; middle and right, the leading areas of mCherry-tagged plasma membrane and histone(red) and ARX-2::GFP knockin (green). Furthest right, the normalized GFP fluorescence distribution plot, as described in (A) to (C). Scale bar, 5 mm.(E and F) Box plots show the distribution of ARX-2::GFP fluorescence intensity ratio between the leading edge and the cytosol of AQR (E) and the percentage ofleading-edge periphery with enriched ARX-2::GFP (F). Enrichment is defined if the GFP intensity in AQR is 1.5 times greater than that in
hyp7. n = 50. The statisticalsignificances are: compared with WT, ***p < 0.001; compared with
wsp-1(
gm324), ##p <0.01 and ###p < 0.001.(G) Triple fluorescence images of AQR. The corresponding normalized fluorescence knockin intensity distribution along the leading edge is shown on the right.Individual puncta of TagRFP::WVE-1 or GFP::WSP-1 are labeled and numbered in representative images and the corresponding plots. The asterisk indicates anoverlapping cluster of TagRFP::WVE-1 and GFP::WSP-1. Hash indicates an autofluorescent particle. Scale bar, 2.5 um.(H) Quantification of the number of puncta of WVE-1 and WSP-1 at the leading edge. The black portion in each bar indicates the overlapped WVE-1 and WSP-1.