Journal: Scientific Reports

Article Title: Evidence for a radiation-responsive ‘p53 gateway’ contributing significantly to the radioresistance of lepidopteran insect cells

doi: 10.1038/s41598-017-18521-5

Figure Lengend Snippet: Ectopic overexpression of Sfp53 induces caspase-3 dependent apoptosis in miR-31 independent manner. ( a ) Sf9 cells were harvested 24 h following transfection with Sfp53 clone. Significant cell death was observed under DIC microscope. Cells were then harvested and immuno-blotted with p53 specific antibody. Cell death/apoptosis was also measured 24 h post transfection by ( b ) caspase-3 activation and ( c ) morphological analysis. Data are mean ± SD of three independent experiments. (*P < 0.001, **P < 0.02) ( d ) Sf-miR-31 showed no change in expression, measured by q-PCR 24 h after Sfp53 clone transfection.

Article Snippet: Cell death was assessed by observing cells under DIC microscope (Axiovert-200 Zeiss inverted DIC microscope, Carl Zeiss, Germany).

Techniques: Over Expression, Transfection, Microscopy, Activation Assay, Expressing

Journal: Nucleic Acids Research

Article Title: A DEAD-box protein acts through RNA to promote HIV-1 Rev-RRE assembly

doi: 10.1093/nar/gkx206

Figure Lengend Snippet: Rev-RRE assembly monitored by single-color TIRF microscopy. ( A ) Schematic of the assembly assay, showing the RRE construct immobilized on a PEG-coated surface by means of biotin–neutravidin attachment. One or more Rev molecules labeled with an A555 fluorophore (green circle) can bind from solution to the immobilized RRE and become fluorescent upon excitation by a totally internally reflected green laser (not shown). ( B ) The left panel shows a representative fluorescence intensity versus time trace (green) recorded in the absence of DDX1 and a corresponding fit to a hidden Markov model (red). The right panel shows a binned histogram of fluorescence intensities compiled from 223 individual intensity-time traces. ( C ) Corresponding data recorded in the presence of 50 nM WT DDX1. The histogram is compiled from 132 individual traces. Other details as in part A. ( D ) Corresponding data recorded in the presence of 50 nM DDX1 (-Rev) mutant. The histogram is compiled from 292 individual traces. Other details as in part A. ( E ) Corresponding data recorded in the presence of 50 nM DDX1 (-RNA) mutant. The histogram is compiled from 142 individual traces. Other details as in part A.

Article Snippet: Single-molecule fluorescence data was collected using a custom built prism-based TIRF microscope (Axiovert 200, Zeiss) under oxygen scavenging conditions, as described ( ).

Techniques: Microscopy, Construct, Labeling, Fluorescence, Mutagenesis