Lf pa0011

Total cell lysates, cytosol and nuclear extract of LECs were prepared and protein blot analysis was performed as described previously [43 (link),52 (link),78 (link),79 (link)]. The membranes were probed with anti-Nrf2 (SC-722, Santa Cruz Biotechnology, Dallas, TX, USA), Anti-Klf9 (ab177158, Abcam^®, Cambridge, MA, USA), Anti-Prdx6 antibody (LF-PA0011, Ab Frontier, South Korea) or β-actin (A2066, Sigma-Aldrich, St. Loius, MO, USA)/Lamin B1 (ab133741, Abcam^®, Cambridge, MA, USA) as internal control to monitor those protein expressions. After secondary antibody (sc-2354 and sc-2768, Santa Cruz Biotechnology, Dallas, TX, USA), protein bands were visualized by incubating the membrane with luminol reagent (sc-2048; Santa Cruz Biotechnology, Dallas, TX, USA) and images were recorded with a FUJIFILM-LAS-4000 luminescent image analyzer (FUJIFILM Medical Systems Inc., Hanover Park, IL, USA).

The total cellular extracts of LECs were prepared in ice-cold radioimmune precipitation buffer (RIPA buffer), and Western analysis was carried out as described previously [7 (link),70 (link),71 (link),72 (link)]. The membranes were probed with anti-Nrf2 (sc-722, Santa Cruz Biotechnology, Dallas, TX, USA), anti-Nrf2 (ab62352, Abcam^®, Cambridge, MA, USA), Anti-Klf9 (ab177158, Abcam^®, Cambridge, MA, USA), Anti-Klf9 (sc-12996, Santa Cruz Biotechnology, Dallas, TX, USA), Anti-Prdx6 antibody (LF-PA0011, Ab Frontier, South Korea), Tubulin (ab44928, Abcam^®, Cambridge, MA, USA) or β-actin (A2066, Sigma-Aldrich, St. Louis, MO, USA) or Lamin B1 (ab133741, Abcam^®, Cambridge, MA, USA) as an internal control to monitor levels of protein expressions. After secondary antibody (sc-2354 and sc-2768, Santa Cruz Biotechnology, Dallas, TX, USA) incubation, protein bands were visualized by incubating the membrane with luminol reagent (sc-2048; Santa Cruz Biotechnology, Dallas, TX, USA). Finally, images (bands) were recorded with a FUJIFILM-LAS-4000 luminescent image analyzer (FUJIFILM Medical Systems Inc., Hanover Park, IL, USA).

Cell lysates of SRA-hLECs and mLECs were prepared in an ice-cold radioimmune precipitation buffer, and protein blot analysis was performed as described previously [78 (link),79 (link),80 (link)]. The membranes were probed with anti-HA (ab 18181 and ab9110, Abcam^®, Cambridge, MA, USA), Anti-Sp1, Anti-Prdx6 antibody (LF-PA0011 and LF-MA0018, Ab Frontier, South Korea), or β-actin (A2066, Sigma-Aldrich, St. Loius, MO, USA) as internal control to monitor those protein expressions. After secondary antibody (sc-2354 and sc-2768, Santa Cruz Biotechnology, Dallas, TX, USA), protein bands were visualized by incubating the membrane with luminol reagent (sc-2048; Santa Cruz, Santa Cruz Biotechnology, Dallas, TX, USA) and images were recorded with a FUJIFILM-LAS-4000 luminescent image analyzer (FUJIFILM Medical Systems Inc., Hanover Park, IL, USA).