Targeting Immune-Related Molecules in Cancer Therapy: A Comprehensive In Vitro Analysis on Patient-Derived Tumor Models

Figure 2

Quantitative gene expression analysis as determined by quantitative PCR (Taqman®). (a) Gene expression changes in HROC cell lines after Indoximod treatment (72 h, monotherapy). (b) Altered gene expression in HROC50 T1 M5 cells after combination with various test substances as stated in material and methods. Reactions were performed in triplicate wells and repeated three times. mRNA levels of target genes were normalized to the housekeeping gene GAPDH. The general expression level of each sample was considered by calculating resulting from the difference between ΔC- ΔC. DMSO-treated cells were used as calibrator. N = 4 independent experiments, mean ± SD, p < 0.05 versus control. Unpaired Student’s t-test. (c) Autophagy was detected by using confocal laser scanning microscopy. Cells were stained with acridine orange (excitation/emission: 500nm/526nm) to visualize acidic lysosomes within the cytoplasma, counterstaining was done with Calcein AM (excitation/emission: 494nm/517nm) as stated in material and methods. In most cases, autophagy was hardly detectable. Original magnification 200x. HROC, Hansestadt Rostock Colon.

(a)

(b)

(c)