Increased miR4485-3p expression in emphysema. ATII cells were isolated from nonsmokers (NS), smokers (SM), and emphysema patients (EM). (A) Sense SncmtRNA and anti-sense ASncmtRNA-1 and ASncmtRNA-2 RNAs levels were normalized to GAPDH and determined by RT-PCR. (B) mir1973 and mir4485-3p expressions were normalized to U6 and analyzed by RT-PCR. (C) A549 cells were transfected with miR-4485-3p or NT for 72 h, and 12S and 16S rRNA expression was determined by RT-PCR and normalized to GAPDH (N = 3 biological replicates). (D) A549 cells were treated with CSE for 4 h, 24 h, 48 h, and 72 h. The expression of 12S and 16S rRNA was determined by RT-PCR and normalized to GAPDH (N = 3 biological replicates). (E) Plasma samples obtained from NS, SM, and COPD (GOLD1–GOLD4) were used to assess the expression of circulating mir-4485-3p (N = 4–9 per group). Results were normalized to U6 and analyzed by RT-PCR. Data are shown as means ± s.e.m (* p < 0.05; ** p < 0.01; *** p < 0.001).