Journal: Cancers

Article Title: Therapeutic and Diagnostic Potential of Folic Acid Receptors and Glycosylphosphatidylinositol (GPI) Transamidase in Prostate Cancer

doi: 10.3390/cancers16112008

Figure Lengend Snippet: Folate receptor 1 expression in healthy PNT2 and prostate cancer cells (LNCaP). Folate receptor 1 (FR1/red) was quantified by qRT-PCR ( A ), Western blot ( B ) and immunofluorescence ( C , D ). The graphs show relative values calculated to healthy control cells. In ( B ), a representative Western blot membrane with the corresponding marker band (black) at 37 kDa is shown. The effective protein load was detected by BioRad-stain-free technology, enabling adjustment of protein loads by quantification of total protein levels. p -values of * 0.05, ** 0.01 and *** 0.001 were labeled with one to three asterisks indicating significances from at least 3 independent experiments. Scalebar 100 µm.

Article Snippet: For all experiments, the two cell lines, LNCaP (Merck, 89110211, prostate cancer human, Rahway, NJ, USA) and PNT2 (Merck, 95012613, healthy epithelial prostate cell), were used.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Membrane, Marker, Staining, Labeling

Journal: Cancers

Article Title: Therapeutic and Diagnostic Potential of Folic Acid Receptors and Glycosylphosphatidylinositol (GPI) Transamidase in Prostate Cancer

doi: 10.3390/cancers16112008

Figure Lengend Snippet: Glycosylphosphatidylinositol transamidase expression and folate receptor 1 localization in prostate cancer cells (LNCaP) and healthy prostate epithelial cells (PNT2). Glycosylphosphatidylinositol transamidase (GPI-T/blue) was quantified by qRT-PCR ( A ), Western blot ( B ) and immunofluorescence ( C , D ) in healthy PNT2 and prostate cancer cells (LNCaP). In ( D ), immunofluorescence against FR1 (red) is additionally shown; white arrows indicate membrane signals. Correlation between FR1-membrane fluorescence intensity and GPI-T fluorescence intensity in healthy PNT2 (gray squares) and LNCaP cancer cells (red circles) with corresponding correlation value Pearson R is shown in ( E ). The graphs ( A – C ) show the relative values calculated for the healthy control cell. ( B ) shows a representative Western blot membrane with the corresponding marker bands at 37 kDa and 50 kDa. The effective protein load was detected by BioRad-stain-free technology, enabling adjustment of protein loads by quantification of total protein levels. p -values of * 0.05, ** 0.01 and *** 0.001 were labeled with one to three asterisks indicating significances from at least 3 independent experiments. Scalebar 100 µm.

Article Snippet: For all experiments, the two cell lines, LNCaP (Merck, 89110211, prostate cancer human, Rahway, NJ, USA) and PNT2 (Merck, 95012613, healthy epithelial prostate cell), were used.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Membrane, Fluorescence, Marker, Staining, Labeling

Journal: Cancers

Article Title: Therapeutic and Diagnostic Potential of Folic Acid Receptors and Glycosylphosphatidylinositol (GPI) Transamidase in Prostate Cancer

doi: 10.3390/cancers16112008

Figure Lengend Snippet: Folate-functionalized lipoplexes for selective transfer of eGFP-plasmid DNA into cancer cells. As a control, eGFP plasmid DNA was transferred into healthy PNT2 and LNCaP cancer cells using unmodified standard lipoplexes. In addition, an increasing concentration of NHS-PEG-FA was used to characterize specific uptake via FR1 examined by eGFP-positive cells using microscopy ( A , B ). In addition, quantification was performed by flow cytometry, which provided information on the changes in transfection efficiency for each cell type ( C ) and the altered specificity between FR1-mediated uptake in PNT2 and LNCaP ( D ). Finally, in co-culture, this specific transfection of standard lipoplexes ( E ) and FA-functionalized lipoplexes ( F ) was further investigated. The healthy cells are shown demarcated in the white frames and could be identified by GPI-T immunofluorescence (weak blue) based on the lower fluorescence intensity compared to LNCaP cells (bright blue). Quantification of enhanced selectivity in co-culture is shown in ( G ). p -values of * 0.05, ** 0.01 and *** 0.001 were labeled with one to three asterisks indicating significances from at least 3 independent experiments. n.s., not significant. Scale bar 100 µm.

Article Snippet: For all experiments, the two cell lines, LNCaP (Merck, 89110211, prostate cancer human, Rahway, NJ, USA) and PNT2 (Merck, 95012613, healthy epithelial prostate cell), were used.

Techniques: Plasmid Preparation, Concentration Assay, Microscopy, Flow Cytometry, Transfection, Co-Culture Assay, Immunofluorescence, Fluorescence, Labeling