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. Author manuscript; available in PMC: 2017 Mar 14.
Published in final edited form as: Cancer Cell. 2016 Mar 14;29(3):367–378. doi: 10.1016/j.ccell.2016.02.012

Figure 2.

Figure 2

Role of PKCι in the LADC TIC phenotype. (A) QPCR of parental and oncosphere cells for PKCι and MMP10. Results expressed as fold parental +/−SEM. n=3. *p<0.05 compared to parental. (B) PKCι knock down (KD) in A549 oncospheres using two RNAi constructs. Immunoblot (top), and QPCR (bottom). QPCR results expressed as fold parental cells +/−SEM, n=3. *p≤ 0.05 compared to parental. (C) Representative micrographs showing effect of PKCι KD on oncosphere growth. Scale bars 50 μm. (D) Oncosphere size expressed as mean diameter (μm) +/−SEM. Oncospheres assessed: A549: NT (63), KD1 (63), KD2 (58); H358: NT (44), KD1 (40), KD2 (39); H23: NT (41), KD1 (33), KD2 (37). *p≤0.05. (E) Single oncosphere cells were assessed for clonal expansion plotted as % expanded +/−SEM. Cells expanded/total cells: A549: NT (55/63), KD1 (7/63), KD2 (7/58); H358: NT (37/44), KD1 (5/40), KD2 (4/39); H23: NT (35/41), KD1 (4/33), KD2 (4/37). *p≤0.0001 compared to NT by Fisher’s exact test. (F) Soft agar growth expressed as colony number +/−SEM, n=6. *p≤ 0.05 compared to NT. (G) Immunoblot of A549 oncosphere cells stably transduced with NT or PKCι RNAi and stably transfected with either empty vector (EV) or vector encoding PKCι (+PKCι). (H) Representative micrographs of PKCι reconstitution on A549 oncosphere growth. Scale bars 50 μm. (I) Oncosphere size expressed as mean diameter (μm) +/−SEM. Number assessed: NT + EV (39), KD + EV (34), KD + PKCι (35). *p≤ 0.05 compared to NT+EV, **p≤0.05 compared to PKCι KD+EV. (J) Soft agar growth plotted as colony number +/−SEM, n=6. *p≤ 0.05 compared to NT+EV, **p≤0.05 compared to PKCι KD+EV. See also Figure S2.

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