Target specific probes

Following hypoxia treatment, NHNE cells were harvested for total RNA extraction. The cells were homogenized in Trizol Reagent (Sigma-Aldrich, St. Louis, MO, USA), and real-time PCR was performed according to the manufacturer's protocol. The primers (β-actin fwd: 5' GCCAACCGC GAGAAGATG-3', rev: 5'ACGGCCAGAGGCGTA CAG-3'; ZO-1 fwd: 5'-TGGTGTCCTACCTAAT TCAACTCA-3', rev: 5'-CGCCAGCTACAAATATTC CAACA-3'; E-cadherin fwd: 5'-ATAGAGAACGCATT GCCACATACA-3', and rev: 5'-TTCTGATCGGTTACC GTGATCA-3') were synthesized, and quantitative reverse transcription-PCR was performed using the TaqMan® universal PCR master mix (Applied biosystems, Foster City, CA, USA). Target-specific probes were purchased from Applied Biosystems. All reactions were performed in triplicate using different batches of NHNE cells. The relative mRNA quantities were determined using the comparative threshold method, and the results were normalized against β-actin.

TaqMan^® MicroRNA Assays (Applied Biosystems, CA, USA) were used to quantitatively detect the levels of miR-301a using target-specific probes (000528, Applied Biosystems) according to the manufacturer’s instructions. RNU6B (001973, Applied Biosystems) was used to normalize the data. The rat specific primers were summarized in Supplemental Table 1. The quantitative RT-PCR reaction was performed on an ABI 7500 Fast Real-Time PCR System.