Fitc conjugated goat anti rat

Immunofluorescence was performed following the protocol described previously11 (link). Primary antibodies that we used include: rabbit anti-mouse LYVE-1 antibody (1:200, AngioBio), rat anti-mouse CD31 (1:100, BD Pharmingen), rabbit anti-phospho-Met (1:400, Cell Signaling), mouse anti-human PCNA (1:100, BD Pharmingen), mouse anti-human vimentin (1:50, Santa Cruz), and goat anti-mouse lectin FITC (1:100, Sigma). Secondary antibodies include: FITC-conjugated goat anti-rat, FITC-conjugated goat anti-rabbit, rhodamine-conjugated goat anti-mouse, Cy3-conjugated goat anti-rabbit, and Alexa Fluor 488 goat anti-rabbit (1:500, all from Jackson Immunoresearch). Fluorescent signals were visualized and digital images were obtained using the LSM-510 confocal microscope (Carl Zeiss). We quantified the images using ImageJ (NIH, Bethesda, MD), measuring the pixel number/10× frame of randomly selected 12 images in each sample. Each group included at least three different samples.