Figure S4.

Figure S4.

(A, B) Pre-abscising zebrafish embryonic cell (A) or a Rab11-null human (HeLa) cell expressing mCh-Rab11 (gray, A and B top panel; cyan in merge, B) fixed and immunostained for γ-tubulin (magenta, A), Pericentrin (magenta, B), and DNA (DAPI, blue, A). Scale bar, 10 μm. Magnified insets of acentrosomal site (a’, a”, and b”) and centrosome (b’). (C) Time-lapse of GFP-FIP3/Rab11 (inverted grays, left; cyan, merge) centrosome region in a pre-abscising human (HeLa) cell expressing DsRed-PACT (inverted grays, left; magenta, merge). Pink arrows, acentrosomal fragments positive for GFP-FIP3/Rab11/DsRed-PACT. Scale bar, 10 μm. (c’) Outline of tracked particles noted by pink arrow. Orange line, centrosome. (D, E) Human (HeLa) pre-abscising cells expressing GFP-FIP3 (cyan, D) or centrin-GFP (cyan, E) were fixed and immunostained for transferrin receptor (fire LUT, D), and cenexin (fire LUT, E). Magnified insets (3×) shown on right (d’, e’) with associated three-dimensional surface plot of intensity. Scale bar, 5 μm. (F) Human GFP-FIP3 (HeLa) cells late in pre-abscission were fixed and immunostained for Pericentrin (fire LUT). The same examples are shown with levels adjusted on right showing that there is some Pericentrin at one of the daughter centrosomes, but it is significantly decreased. (G) Violin plot with median (orange dashed line) and quartiles (dark lines) depicting normalized Pericentrin intensities at centrosomes for n > 20 centrosomes from a representative experiment. Two-tailed t test, ****P < 0.0001. n-values and statistical results detailed in Table S1.