c1si laser scanning confocal microscope (clsm)  (Nikon)

Journal: International Journal of Molecular Sciences

Article Title: Cold Treatment Induces Transient Mitochondrial Fragmentation in Arabidopsis thaliana in a Way that Requires DRP3A but not ELM1 or an ELM1-Like Homologue, ELM2

doi: 10.3390/ijms18102161

Figure Lengend Snippet: ELM2 encodes an ELM1-like protein and GFP-tagged ELM2, like ELM1, localizes to the mitochondrial surface. ( a ) Clustal W alignment of ELM1 and ELM2 amino acids sequences. * depicts the positions of numbers in every ten amino acids. ( b ) Localization of GFP-ELM2 surrounding mitochondria. Arabidopsis cultured cells transiently expressing GFP-ELM2 with a mitochondrial marker MitoTracker were examined by confocal laser scanning microscopy (CLSM). A part of a single cell is shown. Left and Center are separate images obtained with the GFP and MitoTracker, respectively. Right is the merged image. Scale bar, 5 μm, is applicable to the other two figures. Upper right insets are X2 enlarged images.

Article Snippet: A confocal laser scanning microscope (CLSM) (Nikon TE2000-U and C1Si) was used for all microscopic observations of Arabidopsis leaves and cultured cells with fluorescent fusion proteins or stained with fluorescent dyes.

Techniques: Cell Culture, Expressing, Marker, Confocal Laser Scanning Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Cold Treatment Induces Transient Mitochondrial Fragmentation in Arabidopsis thaliana in a Way that Requires DRP3A but not ELM1 or an ELM1-Like Homologue, ELM2

doi: 10.3390/ijms18102161

Figure Lengend Snippet: Disruption of ELM2 does not appear to affect mitochondrial morphology much. ( a ) A T-DNA insertion in the end of the 3rd intron in the elm2 mutant. ( b ) RT-PCR of full length of ELM2 ORF (open reading frame) in the wild-type, elm1-1 , elm2 and elm1-1 elm2 double mutants. ( c ) Comparison of growing phenotypes of wild-type, elm1-1 , elm2 and elm1-1 elm2 double mutants. 30-day-old plants. Scale bar, 5 cm. ( d ) Mitochondrial morphologies in the wild-type, elm1-1 , elm2 and elm1-1 elm2 double mutants. Leaf epidermal cells in 14-day-old plants were observed by confocal laser scanning microscopy. Scale bar, 10 μm, is applicable to the four images. ( e ) Average planar areas of mitochondria of wild type and mutants. ( n > 218 in each of three replications) in each mutant. Error bars show S.E. ** indicates statistical significance at p < 0.01.

Article Snippet: A confocal laser scanning microscope (CLSM) (Nikon TE2000-U and C1Si) was used for all microscopic observations of Arabidopsis leaves and cultured cells with fluorescent fusion proteins or stained with fluorescent dyes.

Techniques: Disruption, Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Comparison, Confocal Laser Scanning Microscopy

Journal: International Journal of Molecular Sciences

Article Title: Cold Treatment Induces Transient Mitochondrial Fragmentation in Arabidopsis thaliana in a Way that Requires DRP3A but not ELM1 or an ELM1-Like Homologue, ELM2

doi: 10.3390/ijms18102161

Figure Lengend Snippet: Heterologous complementation test of mitochondrial morphology in the elm1 mutant by expression of ELM2 . ( a ) Schematic drawing of DNA constructs used in this study. ELM1 , ELM2 and GUS coding sequences are attached between the probable promoter, the 950bp upstream region of ELM1 and the sequence of CaMV35S terminator. ( b ) RT-PCR of the full length of the ELM2 ORF in the wild-type, elm1-1 , and three elm1-1 mutants transformed with ELM1pro:ELM1 , ELM1pro:ELM2 and ELM1pro: GUS respectively. ( c ) Occurrence of elongate mitochondria in leaf epidermal cells in 14-day-old cotyledons from five Arabidopsis lines (wild-type, elm1-1 , and three elm1-1 mutants transformed with ELM1pro:ELM1 , ELM1pro:ELM2 and ELM1pro: GUS ). Occurrence is expressed as the percentage of 40 confocal laser scanning microscopic images obtained from 8 leaves from each line that were judged to have elongated mitochondria (as in the elm1-1 image in ). The experiments were repeated three times independently and the results were averaged. Error bars show S.E.

Article Snippet: A confocal laser scanning microscope (CLSM) (Nikon TE2000-U and C1Si) was used for all microscopic observations of Arabidopsis leaves and cultured cells with fluorescent fusion proteins or stained with fluorescent dyes.

Techniques: Mutagenesis, Expressing, Construct, Sequencing, Reverse Transcription Polymerase Chain Reaction, Transformation Assay