Figure 4. Identification of PKCι-dependent and -independent cells of origin for LADC.

(A and B) Oncosphere number (A) and size (B) of KP and KPI cells grown in non-adherent culture. Results represent mean ±SEM. n=3; *p<0.05 by Student’s t-test.

(C) Micrographs showing the size and morphology of representative KP and KPI oncospheres. Scale bars represent 200 μm.

(D) Abundance of bronchioalveolar stem cells (BASCs) and alveolar type II (AT2) cells in KP and KPI mouse lungs. Data expressed as mean % total lung epithelial cells ±SEM. n=3.

(E) PCR of genomic DNA from KP and KPI BASC and AT2 cells after transduction with Ad-Cre ex vivo. unrec.=unrecombined; rec.=recombined; WT, wild-type. DNA from total lung epithelial cells in the absence of Ad-Cre included as a control.

(F and G) Oncosphere growth of purified BASCs and AT2 cells from KP and KPI mice transduced with Ad-Cre. Oncosphere numbers (F) and size (G) are shown as mean ±SEM. n=3. *p<0.05 compared to KP by Student’s t-test.

(H) Immunofluorescence of a bronchiolar lesion in a KP mouse stained for CCSP (green, upper left panel), SPC (red; upper right panel) or merged image CCSP, SPC and DAPI (blue, lower left panel) six weeks after induction of tumor formation. Lower right panel shows H&E image of same lung region revealing growth of bronchiolar hyperplasia from the affected terminal bronchiole. Scale bars indicate 100 μm.

(I) Immunofluorescence of an alveolar lesion in KPI mouse stained for CCSP (green, upper left panel), SPC (red; upper right panel) or merged image CCSP, SPC and DAPI (blue, lower left panel) six weeks after induction of tumor formation. Lower right panel shows H&E image of same lung region revealing an alveolar lesion without bronchiolar involvement. Scale bars indicate 100 μm.

See also Figure S1.