Research Paper Volume 1, Issue 4 pp 402—411

Figure 3. Overexpression of miR-146a/b suppresses basal and senescence-associated secretion of IL-6 and IL-8 in HCA2 cells. (A) Western blot analyses of TCA-precipitated proteins prepared from CM collected over 24 h from cells infected with the lentivirus backbone (cntrl) or lentiviruses expressing miR-146a (146a) or miR-146b (146b). The blot was analyzed for IL-6, IL-8 and IGFBP3. Equal loading was based on cell number prior to collection of CM and IGFBP3 levels. Proliferating indicates cells described in Figure 2A. The same cells were treated with bleomycin and CM was harvested 11 days later (senescent). (B) IL-6 in CM from the cell populations described in Fig 3A was measured by ELISA. The data are reported as 10^-6 pg per cell per day. (C) RT-PCR analysis of transcript levels of IL-6 and IL-8 in miR-146a/b-overexpressing cells. RNA collected from proliferating cells was used as the control (cntrl).