1. Fig. 4. Induced Zbtb7a in lungs is localized to AMs.
Induced Zbtb7a in lungs is localized to AMs. (A) Representative reverse transcription PCR analysis on total RNA isolated from AMs (lanes 1, 4, 7, and 10), non-AM BAL cells (lanes 2, 5, 8, and 11), and alveolar respiratory epithelial cells (REC; lanes 3, 6, 9, and 12) on day 7 from siRNA lentivirus–transduced and Ab-treated mice (n = 6). bp, base pair. (B) Freshly isolated human AMs (from stable lung transplant recipients; DSA−, autoAb−, and with no documented rejection) and mouse AMs (from C57BL/6) were cultured at 1 × 106 cells per well (33). Surface MHC class I was ligated by anti–H-2Kb and anti-HLA, respectively, in mouse and human AM cultures, and MHC class II on mouse AMs was ligated by anti–I-A/I-E. Isotype-matched controls for MHC Abs and ligation of a non-MHC molecule (keratin) were also evaluated. Zbtb7a expression was normalized to that of Actb and presented as relative expression compared to no Ab–treated AMs. n = 5. Data are presented as means ± SEM, two-tailed unpaired t tests are applied, *P < 0.05 is considered significant, and P values are indicated. (C) BAL cells from five BOS+ lung transplant recipients (P-1 to P-5) and four BOS-free lung transplant recipients (P-6 to P-9) with stable lung function were fractionated into AM and non-AM cells. The lung biopsy specimens were obtained on the same day of BAL fluid collection. Fold induction in ZBTB7A expression compared to the peripheral blood leukocytes for individual lung transplant recipient is plotted as heat map.
Deepak K. Nayak et al., Sci Transl Med 2017;9:eaal1243
Published by AAAS