XB-IMG-173824
Xenbase Image ID: 173824
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Figure S1: Specificity of wdr5 MO and Wdr5 antibody. Related to Figure 1.
(A) Western Blot indicating Wdr5 levels in uninjected controls, wdr5 morphants, and
wdr5 morphants injected with human WDR5 RNA at stage 20. Numbers at the top are
normalized values for Wdr5 intensity based on loading control Gapdh.
(B) Western Blot indicating WDR5 levels in RPE cells using WDR5 antibody, WDR5
antibody mixed with blocking peptide in 1:1 and 1:2 volumetric ratio. A, B represent two
independent protein isolations.
(C) Western Blot indicating Wdr5 levels in uninjected controls, wdr5 morphants, and
wdr5 morphants injected with either human WT-WDR5-GFP RNA, human S91K-WDR5-
3xGFP RNA, or human K7Q-WDR5-3xGFP RNA. Numbers at the top are normalized
values for Wdr5 intensity based on loading control Gapdh.
(D-J) Percentage of embryos with present (magenta) or reduced (green) cilia driven
epidermal flow.
(D) Experimental conditions include uninjected controls and scrambled MO
(E) Experimental conditions include uninjected controls, wdr5 MO and wdr5 MO +
human WDR5-GFP.
(F) Experimental conditions include uninjected controls and second wdr5 MO.
(G) Experimental conditions include uninjected controls and wdr5 CRISPR.
(H) Experimental conditions include uninjected controls, wdr5 MO, wdr5 MO + human
WDR5 (K7Q)-3xGFP RNA, and human WDR5 (K7Q)-3xGFP RNA.
(I) Experimental conditions include uninjected controls, wdr5 MO, wdr5 MO + human
WT-WDR5 3xGFP RNA, and wdr5 MO + human WDR5 (26-334)-3xGFP RNA.
(J) Experimental conditions include uninjected controls, injected with wdr5 MO, wdr5 MO
+ human WT WDR5 RNA, and wdr5 MO + human mutant S91K WDR5 RNA.
n = number of embryos. ★★★ indicate statistical significance at P < 0.0005
(K) X. tropicalis epidermal MCCs in controls and CRIPSR injected embryos marked with
anti-acetylated α-tubulin (green) for cilia and phalloidin (magenta) for actin.
(L) Transcriptional profile with absolute number of transcripts per embryo of X. tropicalis
starting at single cell up to stage 28. Image published in: Kulkarni SS et al. (2018) Copyright © 2018. Image reproduced with permission of the Publisher, Elsevier B. V.
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