Quantitative real-time PCR (qPCR)
For the quantification of the overall gene expression of twoPTPRT downstream target genes, a qPCR was performed in a 7500
Fast Real-Time PCR System (Applied Biosystems) and using tumor cDNA from
the carriers, the TaqMan Gene Expression Master Mix (Applied Biosystems)
and specific TaqMan probes designed by Thermo Fisher Scientific
(BCL-XL assay Hs00236329_m1 and SOCS3 assay
Hs02330328_s1). PSMB4 was used as a housekeeping gene whose
levels served as a reference (assay Hs00160598_m1), and a pool of
healthy colon cDNAs was used as a control. All samples were analyzed in
triplicates, and the quantification of the relative target gene
expression was calculated as 2-ΔΔCt. The standard
deviation was calculated for each sample.