FIGURE

Figure 3

ID

ZDB-FIG-221020-18

Publication

Revathidevi et al., 2022 - AMBRA1 p.Gln30Arg Mutation, Identified in a Cowden Syndrome Family, Exhibits Hyperproliferative Potential in hTERT-RPE1 Cells

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Figure 3

AMBRA1 Q30R mutant hTERT-RPE1 cells exhibited abnormal primary ciliogenesis. (A) Immunocytochemistry images of cells stained with Arl13B (ADP Ribosylation Factor Like GTPase 13B), a primary cilia marker (stained green), and DAPI (4′,6-diamidino-2-phenylindole), a nucleus marker (stained blue). White scale bar = 25 μm (B) Bar graph representing the percentage of ciliated cells in each AMBRA1 genotype in nutrient-sufficient condition. Statistical significance was determined by ANOVA. (C) Bar graph representing the percentage of ciliated cells in each AMBRA1 genotype under starvation. Statistical significance was determined by ANOVA. (D) Representative immunocytochemistry images of AMBRA1^{Q30R A/G} mutant cells stained with Arl13B and DAPI, showing abnormal ciliary structures (curled—top; double cilia—bottom left; spiral—bottom right). Magenta scale bar = 15 μm (E) ambra1a zebrafish CRISPANT showing a situs inversus phenotype. The arrow indicates the reverse orientation of the gall bladder in ambra1a knockout zebrafish (top) and its normal orientation in wildtype zebrafish (bottom). (F) Bar graph representing the percentage of zebrafish showing situs inversus phenotype. Statistical significance was determined using Fisher’s exact test.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT + CRISPR6-ambra1a + CRISPR7-ambra1a + CRISPR8-ambra1a + CRISPR9-ambra1a
Knockdown Reagents:	CRISPR6-ambra1a CRISPR7-ambra1a CRISPR8-ambra1a CRISPR9-ambra1a
Observed In:	gall bladder gall bladder determination of left/right symmetry whole organism right side
Stage:	Protruding-mouth

Phenotype Detail

Acknowledgments

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