Journal: Cellular Oncology (Dordrecht)

Article Title: Alterations of monocarboxylate transporter densities during hypoxia in brain and breast tumour cells

doi: 10.1007/s13402-012-0081-9

Figure Lengend Snippet: Electron micrographs of membrane area. a Illustration of the tumour cells ( grey ), gold particles ( yellow ) and membrane area ( green ). b Low magnification transmission electron micrograph from T-47D cells. c High magnification from ( b ) green square , cell membrane region. d Low magnification from T98G cells. e High magnification from ( d ) green square . The morphology is characteristic of tumour cells with nuclear atypia. The area between plasma membranes of two cells is covered with microvillous processes extended from the cell surface. Blue area , cell body; Blue lines , cell membranes; Red lines , nuclear membranes; Green square , membrane area; Scale bars , 5 μm in B and D, 500 nm in C and E; insets, 50 nm in C and E

Article Snippet: Human T-47D breast cancer cells and human glioblastoma T98G cells (purchased from ATCC, LGC Standards AB, SE-501 17 Boras, Sweden) were grown as monolayer cultures in RPMI (Roswell Park Memorial Institute) 1640 medium (JRH Biosciences, Lenexa, KS, USA), supplemented with 10 % fetal calf serum (Gibco, Paisley, UK), 2 mM L-glutamine (SIGMA, St Louis, MO, USA), 200 units l −1 insulin (SIGMA), and 1 % penicillin/streptomycin (Gibco) at 37 °C in air containing 5 % CO 2 .

Techniques: Membrane, Transmission Assay, Clinical Proteomics

Journal: Cellular Oncology (Dordrecht)

Article Title: Alterations of monocarboxylate transporter densities during hypoxia in brain and breast tumour cells

doi: 10.1007/s13402-012-0081-9

Figure Lengend Snippet: Electronic micrographs from T-47D and T98G cells with different MCTs. In electron micrographs, gold particles signalled immunoreactivity to MCT. To quantify the labelling of different transporter subtypes, the area between two cells creating a border region was calculated and gold particles ( red arrows ) situated in the region, including 25 nm on each side of the membranes (approximately the same distance as the lateral resolution of the immunogold method (Bergersen et al. 2008)), were marked by ImageJ software. The density of transporters’ immunoreactivity in the region containing microvillous processes of the surface membrane was calculated as the number of gold particles divided by the area. Red arrows , gold particles; Scale bars, 500 nm; insets, 100 nm

Article Snippet: Human T-47D breast cancer cells and human glioblastoma T98G cells (purchased from ATCC, LGC Standards AB, SE-501 17 Boras, Sweden) were grown as monolayer cultures in RPMI (Roswell Park Memorial Institute) 1640 medium (JRH Biosciences, Lenexa, KS, USA), supplemented with 10 % fetal calf serum (Gibco, Paisley, UK), 2 mM L-glutamine (SIGMA, St Louis, MO, USA), 200 units l −1 insulin (SIGMA), and 1 % penicillin/streptomycin (Gibco) at 37 °C in air containing 5 % CO 2 .

Techniques: Software, Membrane

Journal: Cellular Oncology (Dordrecht)

Article Title: Alterations of monocarboxylate transporter densities during hypoxia in brain and breast tumour cells

doi: 10.1007/s13402-012-0081-9

Figure Lengend Snippet: T-47D and T98G cells were grown either in 20 % or 4 % oxygen. The medium was changed on day 2. a The change in pericellular oxygen concentration as cell number increases between reseedings in cells flasks cultured with 4 % oxygen in the gas phase. Results from one representative experiment are shown. b T-47D cells and T98G cells were seeded at numbers resulting in full but not confluent flasks at day 4 for the MCT analysis. In addition measurements of lactate, glucose and pH were done in flasks with the same number of T98G cell as T-47D cells. c The cellular mass calculated from the cell volume and cell numbers from panel b. Panels d – f show the data after the medium change. d Glucose concentration ( black square shows measurement of medium). e Lactate concentration ( black square shows measurement of medium). f pH. Error bars show standard error of the mean from two parallel flasks

Article Snippet: Human T-47D breast cancer cells and human glioblastoma T98G cells (purchased from ATCC, LGC Standards AB, SE-501 17 Boras, Sweden) were grown as monolayer cultures in RPMI (Roswell Park Memorial Institute) 1640 medium (JRH Biosciences, Lenexa, KS, USA), supplemented with 10 % fetal calf serum (Gibco, Paisley, UK), 2 mM L-glutamine (SIGMA, St Louis, MO, USA), 200 units l −1 insulin (SIGMA), and 1 % penicillin/streptomycin (Gibco) at 37 °C in air containing 5 % CO 2 .

Techniques: Concentration Assay, Cell Culture

Journal: Cellular Oncology (Dordrecht)

Article Title: Alterations of monocarboxylate transporter densities during hypoxia in brain and breast tumour cells

doi: 10.1007/s13402-012-0081-9

Figure Lengend Snippet: Antibody specificity and MCT protein quantification. a Immunoblots from T98G tumour cells and T-47D cancer cells gave for all antibodies single bands at a molecular weight of approximately 49–58 KD for MCT1 M (55 KD), MCT2 G (58 KD), MCT2 H (60 KD), MCT4 A (49 KD) and MCT4 H (52 KD). The results are consistent with the molecular mass of the proteins and the descriptions from the antibody suppliers. b The immunoreactive bands show the expression of each MCT in hypoxia and normoxia, in T-47D and T98G tumour cells. c The level of MCT1, MCT4 and MCT2 increased in T-47D cells after hypoxic cultivation, whereas a significant MCT2 expression decrease was detected in hypoxic T98G cells. Error bars show standard error of the mean. The asterisk indicates a statistically significant difference ( P < 0.05) between hypoxia and normoxia

Article Snippet: Human T-47D breast cancer cells and human glioblastoma T98G cells (purchased from ATCC, LGC Standards AB, SE-501 17 Boras, Sweden) were grown as monolayer cultures in RPMI (Roswell Park Memorial Institute) 1640 medium (JRH Biosciences, Lenexa, KS, USA), supplemented with 10 % fetal calf serum (Gibco, Paisley, UK), 2 mM L-glutamine (SIGMA, St Louis, MO, USA), 200 units l −1 insulin (SIGMA), and 1 % penicillin/streptomycin (Gibco) at 37 °C in air containing 5 % CO 2 .

Techniques: Western Blot, Molecular Weight, Expressing

Journal: Cellular Oncology (Dordrecht)

Article Title: Alterations of monocarboxylate transporter densities during hypoxia in brain and breast tumour cells

doi: 10.1007/s13402-012-0081-9

Figure Lengend Snippet: The density of MCTs on plasma membranes and cytoplasm. The density of MCTs on the membranes of hypoxic tumour cells and normoxic cells were significantly higher than that on cytoplasm areas without mitochondria areas. a–c The MCT1 and MCT4 density on membranes of hypoxic cells was significantly higher than that of normoxic cells in both T-47D and T98G cells. d–e The MCT2 density of T-47D hypoxic cells was significantly higher than that of normoxic cells. For T98G cells, it is the reverse. Error bars show standard error of the mean. Asterisks indicate statistically significant differences ( p < 0.05) between hypoxia and normoxia

Article Snippet: Human T-47D breast cancer cells and human glioblastoma T98G cells (purchased from ATCC, LGC Standards AB, SE-501 17 Boras, Sweden) were grown as monolayer cultures in RPMI (Roswell Park Memorial Institute) 1640 medium (JRH Biosciences, Lenexa, KS, USA), supplemented with 10 % fetal calf serum (Gibco, Paisley, UK), 2 mM L-glutamine (SIGMA, St Louis, MO, USA), 200 units l −1 insulin (SIGMA), and 1 % penicillin/streptomycin (Gibco) at 37 °C in air containing 5 % CO 2 .

Techniques: Clinical Proteomics