Using a TMA (tumour micro array) slide, total and phosphorylated AMPK were analysed by immunohistochemistry, as well as phosphorylated ACC expression in normal and PTC samples from the same patients.
The nuclei micro-array slides were pretreated as described previously [ 11].
By performing FISH on nuclei micro-array slides, many cases could be finished simultaneously on a single slide with high comparability and little variance, making it more simple, rapid and inexpensive to be carried out.
FISH for detection of t(11 14) translocation was performed on nuclei micro-array slides using commercially available LSI IgH/CCND1 dual-color, dual-fusion translocation probe, which was a mixture of the LSI IgH probe labeled with Spectrum Green and the LSI CCND1 probe labeled with Spectrum Orange (Vysis).
The indicators collate 10 predictors for each selected feature and represent their consensus with a colour, akin to a micro-array slide (Fig. 1a ).
After hybridization, the micro-array slide assembly was washed sequentially in 1×SSC-0.2% SDS, 0.1×SSC-0.2% SDS and 0.1×SSC for 5 min at room temperature and spun dry in a low-speed centrifuge.
Immunohistochemistry was performed using antibodies to PLK1, MDM2 and Ki67 on Tissue Micro-Array (TMA) slides of a cohort of 215 primary breast cancers.
Unstained human tissue micro-array (TMA) slides BR451, BR480, BR481, BR701, BR721, BR722, BR801, BR1001, and BR1003 (with no overlapping cases) were purchased from US Biomax (Rockville, MD, USA).
Immunohistochemistry was performed using antibodies to phospho-AMPK (pAMPK), phospho-Acetyl Co-A Carboxylase (pACC) an established target for AMPK, HER2, ERα, and Ki67 on Tissue Micro-Array (TMA) slides of two cohorts of 117 and 237 primary breast cancers.
Afterwards, slides were washed according to the protocol and scanned with a G2505B Micro Array Scanner (Agilent Technologies).
