Oct4A, nestin, and NeuN are expressed in lung cancer cell lines. (A) PCR product of quantitative PCR on lung cancer cell lines using primer set 2 showing Oct4A expression. (B-a) Western blot analysis using antibody recognizing Oct4A (40 kd). (B-b) Western blot using only secondary antibody (C) Western blot analysis of Oct4 showing siRNA knockdown of Oct4. (D) Immunoflourscent imaging of H1299 cells showing nuclear expression of Oct4 (black arrows show positive cells). (E) RT-PCR demonstrating nestin expression in lung cancer cell lines shown in Figure 1A (lanes 1–6). Lane 8, is actin control, lane 7 negative control. (F) Western blot analysis showing nestin expression in lung cancer cell lines. (G) Immunoflourscent imaging of H1299 cells showing expression of nestin in cytoplasmic filaments (40x). (H) Immunoflourescent imaging and corresponding phase contrast image of H1299 cells showing nestin is expressed only in a subset of cells (20x). White arrows show immunopositive cells. (I) Western blot analysis showing the expression of NeuN in lung cancer cell lines. (J) Immunoflourescent imaging and corresponding phase contrast image showing nuclear expression of NeuN occurs in a subset of cells (20x). (K) FACS analysis for CD44 expression on H1299 cells. (a) control cells stained with phycoerythrin (PE) secondary antibody are without fluorescence. (b) Greater than 99% of cells stained with anti-CD44 and PE secondary demonstrated fluorescence.