TOLLIP expression in nasal and alveolar epithelium. TOLLIP expression in nasal and alveolar epithelium. (A) T84 cells were plated at two different cell densities: 5×105 per well (lanes 1, 2); 2×106, (lanes 3, 4). Lane 5 represents a negative control without the reverse transcriptase. GAPDH was used as a housekeeping gene. (B) TOLLIP expression was quantified in primary nasal and alveolar epithelium. *p<0.05 by Mann-Whitney U test. (C and D) Cell lines were infected with Staphylococcus aureus strain Newman. RNA extraction was performed followed by RT-PCR. Panel C shows RPMI 2650 cells—and panel D A549 cells—infected with S. aureus. Lanes: (1) positive control for TOLLIP from cell line T84; (2 and 3) unstimulated; (4 and 5) cells with S. aureus at 1.1×105 cfu/mL; (6 and 7) cells with S. aureus at 1.6×105 cfu/mL. GAPDH was used as a housekeeping gene. Band size for TOLLIP 347 bp and for GAPDH 727 bp (TOLLIP, Toll-interacting protein; RT-PCR, reverse transcriptase PCR). Olga Lucia Moncayo-Nieto et al. BMJ Open Resp Res 2014;1:e000046 ©2014 by British Thoracic Society