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. 2017 Jan;56(1):131–144. doi: 10.1165/rcmb.2016-0166OC

Figure 4.

Figure 4.

ETS1 knockdown attenuates increases in miR-27a and ET-1 levels and decreases in PPARγ expression in hemin-treated HPAECs. (A) Whole lung homogenates were collected from SS and AA mice. Lung ETS1 levels were measured with qRT-PCR and expressed relative to lung 9S mRNA. *P < 0.05 versus AA (n = 6). (B) HPAECs were treated with dimethyl sulfoxide vehicle (CON) or hemin (HEM, 5 µM) for 72 hours. Mean HPAEC ETS1 levels were measured with qRT-PCR. Each bar represents the mean ETS1 level ± SE relative to 9S expressed as fold change versus CON. *P < 0.05 versus CON (n = 5). (C–E) HPAECs were treated with 10 nM scrambled (CON) or small interfering RNA duplexes to ETS1 for 72 hours. HPAEC mRNA or protein was then isolated. Each bar represents the mean ± SE (C) ETS1 or (D) PPARγ relative to 9S in the same sample expressed as fold change versus CON. *P < 0.05 versus CON (n = 3). Western blotting was used to detect (C) ETS1 and (D) PPARγ protein levels. Representative blots depicting ETS1 or PPARγ protein in siETS1-treated HPAECs are shown. (E–G) HPAECs were treated with 10 nM scrambled (CON) or small interfering RNA duplexes to ETS1 for 6 hours and then treated with dimethyl sulfoxide vehicle (CON) or hemin (HEM, 5 µM) for 72 hours. MicroRNAs (miRNA) or mRNA were isolated and subjected to qRT-PCR analysis for miR-27a, PPARγ, ET-1, and RNU6B or 9S. Each bar represents the mean ± SE (E) miR-27a, (F) PPARγ, or (G) ET-1 relative to RNU6B or 9S expressed as fold change versus CON. *P < 0.05 versus CON; +P < 0.05 versus HEM (n = 3–6). ETS1, v-ets avian erythroblastosis virus E26 oncogene homolog 1; SCR, scrambled small interfering RNA; siETS1, ETS1 small interfering RNA.