Fig 1.
Real-Time PCR quantification of mRNA.
Real-time PCR showing relative expression levels of SSTR2 (A) and NCL (B) in a panel of human cell lines displayed on a logarithmic and a non-logarithmic scale, respectively. Expression levels were normalized to GAPDH and ACTB as housekeeping genes. We defined a common reference value (REF; see Materials and methods), to allow for comparison of expression levels between the two genes. The average level and SEM for each cell line were calculated from three wells.
Fig 2.
Western blot detection of NCL.
(A) Western blot of extracts from different cell lines were first probed with anti-NCL antibody and afterwards reprobed with anti-GAPDH antibody for equal loading control. (B) Quantification of NCL expression relative to the expression of GAPDH and normalized to the mean value of the normal epithelial breast cell lines MCF-10A and 184A1.
Fig 3.
(A) The two NSCLC cell lines H1299 and A549 were grown as adherent cells or spheres for six days. (B) The number of lung cancer spheres after six days at different number of seeded cells. Mean and SEM from 3–4 plates. (C) Diameter of A549 and H1299 lung cancer spheres. Mean and SEM of 3–4 plates. The number of H1299 spheres (D) and A549 spheres (E) correlated with cell viability. SEM for sphere count and cell viability from 3–4 plates plotted as vertical and horizontal error lines, respectively. T-test depicts difference between A549 and H1299 at identical number og seeded cells p-value <0.05; *, <0.01; **; <0.0001; ****.
Fig 4.
Uptake of 57Co labeled DOTATATE in the NSCLC cell line H1299.
The subcellular distribution of 57Co-DOTA-AS1411 in H1299 adherent cells (A) and in H1299 spheres (B) as a function of increasing incubation time (1–23 h). In each case a blocking experiment was performed with 1000-fold excess AS1411 (23h block). Mean uptake (mBq/cell) is expressed as mean ± SEM from triplicate experiments.