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SAS institute
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Image Search Results
Journal: eLife
Article Title: De novo centriole formation in human cells is error-prone and does not require SAS-6 self-assembly
doi: 10.7554/eLife.10586
Figure Lengend Snippet: ( A, B ) Wild-type (WT) or clone #2 acentrosomal ( p53 -/- ; SAS-6 -/- ) RPE1 cells were stained with indicated antibodies during interphase ( A ) and mitosis ( B ). DNA (DAPI, blue). ( C, D ) Clone #2 acentrosomal cells were infected with lentiviruses, and induced to express indicated SAS-6 constructs for 16h ( C ) or 3 days ( D ). The ability of each SAS-6 mutant in rescuing de novo centriole ( C ) or centrosome ( D ) formation was examined with indicated antibodies. HA staining marks SAS-6. DOI: http://dx.doi.org/10.7554/eLife.10586.005
Article Snippet: De novo centrioles formed in the absence of SAS-6 self-oligomerization can ciliate and duplicate. ( A ) Isogenic SAS-6 -/- ; p53 -/- cells carrying indicated SAS-6 constructs (see ‘Methods’) were arrested in S phase and induced to express indicated SAS-6 mutants for 16 hr and then immunostained with indicated antibodies to visualize
Techniques: Staining, Infection, Construct, Mutagenesis
Journal: eLife
Article Title: De novo centriole formation in human cells is error-prone and does not require SAS-6 self-assembly
doi: 10.7554/eLife.10586
Figure Lengend Snippet: ( A ) Isogenic SAS-6 -/- ; p53 -/- cells carrying indicated SAS-6 constructs (see ‘Methods’) were arrested in S phase and induced to express indicated SAS-6 mutants for 16 hr and then immunostained with indicated antibodies to visualize de novo centrioles (centrin, green; SAS-6/HA, red). DNA (DAPI, blue). ( B ) Isogenic SAS-6 -/- ; p53 -/- cells carrying indicated SAS-6 constructs (see ‘Methods’) were induced to express indicated SAS-6 mutants for 3 days, arrested in G1 for additional 36 hr, and then processed for immunofluorescence microscopy to visualize cilia (GT335, green) and distal appendage (CEP164, red). DNA (DAPI, blue). ( C ) Isogenic SAS-6 -/- ; p53 -/- cells carrying indicated SAS-6 constructs (see ‘Methods’) were induced to express indicated SAS-6 mutants for 3 days, and then processed for BrdU pulse-chase before fixation for immunofluorescence. Centriole duplication was revealed with indicated antibodies (centrin, green; daughter centriole marker, STIL, red; PCM marker, γ-tubulin, blue). S-phase cells labeled with BrdU were shown (blue). ( D ) Quantification of the centriole duplication efficiency from ( C ). S-phase cells were identified (BrdU+) and their centrioles were analyzed by immunofluorescence with centrin, STIL, and γ-tubulin antibodies. Error bars represent standard error of the mean (SEM); n > 150, N = 3. DOI: http://dx.doi.org/10.7554/eLife.10586.006
Article Snippet: De novo centrioles formed in the absence of SAS-6 self-oligomerization can ciliate and duplicate. ( A ) Isogenic SAS-6 -/- ; p53 -/- cells carrying indicated SAS-6 constructs (see ‘Methods’) were arrested in S phase and induced to express indicated SAS-6 mutants for 16 hr and then immunostained with indicated antibodies to visualize
Techniques: Construct, Immunofluorescence, Microscopy, Pulse Chase, Marker, Labeling