Zen 3.3 blue software

2D epifluorescence images were proceeded using ZEN 3.3 blue software (Zeiss). 3D acquired from confocal and multiphoton microscopes were proceeded using NIS-Elements software package. (5.30.03 Nikon Instruments Inc., Nikon Europe B.V., Amsterdam, Netherland). 3D Images acquired from light sheet were proceeded using Imaris software (9.1.2, Oxford Instruments, Abingdon, UK).
Data were analyzed using the GraphPad Prism v5.0 statistical software package (GraphPad Software, La Jolla, CA, USA). Since the data were non normally distributed (normality tested using Shapiro-Wilk test) the non-parametric Mann-Whitney test has been chosen. Significance is depicted (** p<0.01, **** p<0.0001).

Ad5mK8-nlsCre (VVC-Li-535; University of Iowa Viral Vector Core) was diluted in DMEM with CaCl2 (10 mM) and introduced into the fourth mammary ducts at a titer of ~1.92 × 10⁷ pfu/duct using 100 µL Nanofil syringes (World Precision Instruments) fitted with 35 G Nanofil needles (World Precision Instruments). Live animal and organ imaging was performed using an IVIS Spectrum imaging system (Perkin Elmer). Imaging of lung metastases was performed on a Zeiss Axio Observer.Z1 microscope with Zeiss ZEN 3.3 Blue software.