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The specificity of primers uniquely amplifying the four STAT3 splice variants was validated using mixed isoforms as template in PCR. An annealing temperature of 66°C was required to exclude S-α and S-β amplifications from the ΔS-α and ΔS-β reactions, respectively. DNA ladder is exACTGene 1 Kb Plus DNA Ladder (Fisher Scientific International Inc., Waltham, MA).
