image analyzer software package  (Alpha Innotech)

Journal: Cell reports

Article Title: COX7A2L is a mitochondrial complex III-binding protein that stabilizes the III 2 +IV supercomplex without affecting respirasome formation

doi: 10.1016/j.celrep.2016.07.081

Figure Lengend Snippet: (A) Confocal microscopy of 143B cells transiently transfected with the COX7A2L-GFP construct and with the empty-GFP vector as a control. Upper images show the GFP signal. Middle images show the mitochondrial network using an antibody against the ATPase a subunit. Lower images show the overlay between the two signals. (B) BN-PAGE and western-blot analyses of control 143B cells transiently transfected both with the COX7A2L-GFP or COX7A2L-MYC-DDK constructs, and with their corresponding empty vectors. Membranes were incubated with antibodies raised against COX7A2L and the indicated OXPHOS subunits. (C) Densitometric analysis of the MRC complexes and SCs in 143B cells transfected with both COX7A2L- tagged constructs. The optical densities of immunoreactive bands that had not reached saturation levels were measured with the ChemiDoc™ MP Image Analyzer software package (Biorad). The antibody signals within the same structures were quantified; the mean values were normalized by CII and expressed as percentages of the cells transfected with the empty vectors (horizontal bar). Values represent the mean ± SD from four independent experiments. I+III2+IVn, SC containing CI, CIII and CIV. III2+IV, SC containing CIII and CIV. III2, complex III dimer. IV, complex IV. IV2, complex IV dimer. II, complex II. See also Figure S3.

Article Snippet: The optical densities of immunoreactive bands that had not reached saturation levels were measured with the ChemiDoc™ MP Image Analyzer software package (Biorad).

Techniques: Confocal Microscopy, Transfection, Construct, Plasmid Preparation, Western Blot, Incubation, Software