Histological Analysis of Tumor Samples

Histological analysis of primary tumors was performed for a randomly selected subset of CTRL and miR-127^PD tumors. Immunohistochemistry was performed as described (21 (link)). Anti-Ki-67 (1:500) (Cell Signaling Technologies, Cat# 9027), anti-cleaved Caspase 3 [Asp175] (1:500) (Cell Signaling Technologies, Cat# 9661), anti-human mitochondria [113-1] (1:1000) (Abcam, Cat# ab92824), anti-CyclinD1 (Novus Biologicals, Cat# NB100-79920) (1:100), anti-Vimentin (Gentex, Cat# GTX100619) (1:300), anti-Twist1/2 (Genetex, Cat# GTX127310) (1:200), and anti-N-Cadherin (Epitomics, Cat# 2447-1) (1:100), primary antibodies were used. An internal negative control (no primary antibody) was included with each trial. Images of 4 – 6 randomized fields of view per animal were captured then quantified using H DAB color deconvolution software in Image J Fiji (National Institutes of Health, RRID: SCR_002285). For analysis of tumor cell infiltration into lymph node tissue (n = 14), Image J software was used to quantify the total area of the lymph node compared to the total area comprised of tumor cells.