Sirna duplex oligonucleotides

siRNA duplex oligonucleotides against p63 and JAM-A were synthesized by Santa Cruz Biotechnology (Texas, USA). siRNA duplex oligonucleotides against ΔNp63 and GATA-3 were synthesized by Invitrogen (Carlsbad, CA). The sequences were as follows: siRNA of p63 (sense: 5′-GGAAUGACUUCAACUUUGA-3′; antisense: 5′-UCAAAGUUGAAGUCAUUCC-3′), siRNA of JAM-A (sense 5′-UUCGAGUAAGAAGG UGAUUUATT-3′), siRNA of ΔNp63 (sense: 5′-ACAAUGCCCAGACUCAAUU-3′; antisense: 5′-AAUUGAGUCUGGGCAUUGU-3′), siRNA of GATA-3 (sense: 5′-GAGAAA GAGUGCCUCAAGU-3′; antisense: 5′-ACUUGAGGCACUCUUUCUC-3′). Detroit 562 cells and primary cancer cells were transfected with siRNAs of p63, ΔNp63, GATA-3 and JAM-A using Lipofectamine^™ RNAiMAX Reagent (Invitrogen) at 24 h after plating.

siRNA duplex oligonucleotides against LSR (forward sense 5′-CCCACGCAACCCAUCGUCAUCUGGA-3′, reverse sense 5′-UCCAGAUGACGAUGGGUUGCGUGGG-3′) were synthesized by Thermo Fisher Scientific (Waltham, MA). siRNA duplex oligonucleotides against AMOT (sc-72489), YAP (sc-38637) and AREG (sc-39412) were synthesized by SantaCruz Biotechnology, Inc. (Santa Cruz, CA). A scrambled siRNA sequence (BLOCK-iT Alexa Fluor fluorescent, Invitrogen) was employed as control siRNA. At 24 h after plating, Sawano cells were transfected with 100 nM siRNAs of LSR, AREG, YAP and AMOT using Lipofectamine^TM RNAiMAX Reagent (Invitrogen). Some cells were transfected with 100 nM siRNA of LSR with or without 10 μM dobutamine, 500 μM 2-DG and glucose-free medium (Glucose free MEM, Sigma-Aldrich).