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. 2017 Apr 12;8(26):42817–42835. doi: 10.18632/oncotarget.17069

Figure 2. PON1 is located in the cytosolic fraction and is differentially expressed in various lung normal and tumor cell lines.

Figure 2

(A) PON1 protein expression in different lung normal (L132) or cancer cell lines (A549, H460, H1299, H358) including another normal cell line (HEK-293). (B) Western blot analysis of nuclear and cytosolic cell extracts (50 μg of each fraction). (C) Live A549 and A549-shPON1 cells stained with reduced FITC (pseudocolored in green) and DAPI. Cells were analyzed by confocal microscopy. Scale bars = 40 μm or 2 μm as indicated. (D, E) Stable overexpression and knockdown of PON1 by lentiviral-induced systems were examined by Western blotting. In PON1 overexpression system (H460- and H1299-PON1), two isoforms of stable PON1 overexpression (#1 and #2, respectively) differ in lentiviral particles used for transfection. (F) Immunoprecipitation of cells with anti-human PON1 antibody. Representative examples of three independent experiments are shown. All experiments were conducted at least in triplicate and biologically repeated at least twice. Statistical analysis was performed using unpaired Student's t test.