Journal: Frontiers in Molecular Neuroscience

Article Title: Enhancement of Astroglial Aerobic Glycolysis by Extracellular Lactate-Mediated Increase in cAMP

doi: 10.3389/fnmol.2018.00148

Figure Lengend Snippet: Inhibition of AC reduces the L -lactate- and 3Cl-5OH-BA-induced increase in [cAMP] i and [lactate] i (A,B , panels i) Mean time-courses of FRET ratio signal changes normalized to the maximum signal change for (A,i) Epac1-camps upon addition of 20 mM L -lactate and (B,i) Laconic upon addition of 0.5 mM 3Cl-5OH-BA (black lines) in the absence (white circles) and presence (black circles) of 100 μM DDA, an inhibitor of AC. Each data point represents mean ± s.e.m. ( A,B , panels ii) Mean relative changes in FRET ratio (Rel. ΔFRET ratio) upon L -lactate (A) and (B) 3Cl-5OH-BA stimulation in the absence and presence of DDA. Relative ΔFRET values (%) were calculated by dividing individual ΔFRET values with the average ΔFRET value upon L -lactate or 3Cl-5OH-BA stimulation. Note that the inhibition of AC by DDA causes ∼50 % reduction in L -lactate-induced increase in [cAMP] i in astrocytes and a ∼30–60% reduction in 3Cl-5OH-BA-induced increase in [lactate] i in astrocytes, 3T3-L1 and BT474 cells. In BT474 cells the application of 3Cl-5OH-BA initiated a transient reduction in [lactate] i that was diminished in the presence of DDA. Numbers adjacent to black bars represent number of cells. Data are in the format of the mean ± s.e.m ( ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001). Data for every set of experiment was acquired from at least two different animals.

Article Snippet: BT474 cancer cells (BT-474 Clone 5; ATCC ® CRL-3247 TM ; ATCC-LGC Standards) were grown in Hybri-Care medium (ATCC ® 46-X TM ; ATCC-LGC Standards) supplemented with 1.5 g/L sodium bicarbonate and 10% fetal bovine serum.

Techniques: Inhibition

Journal: Breast Cancer : Targets and Therapy

Article Title: Anticarcinogenic Effects of Odorant Substances Citral, Citrathal R and Cyclovertal on Breast Cancer in vitro

doi: 10.2147/BCTT.S322619

Figure Lengend Snippet: MTT-assay. ( A ) MTT-assay T-47D cells, ( B ) MTT-assay BT474 cells, ( C ) MTT-assay MDA-MB-231 cells.

Article Snippet: BT474 cells (#300131) and T-47D cells (#300353) were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany), MDA-MB-231 cells (#92020424) from Sigma-Aldrich (Taufkirchen, Germany).

Techniques: MTT Assay

Journal: Breast Cancer : Targets and Therapy

Article Title: Anticarcinogenic Effects of Odorant Substances Citral, Citrathal R and Cyclovertal on Breast Cancer in vitro

doi: 10.2147/BCTT.S322619

Figure Lengend Snippet: Ki67 in T-47D, MDA-MB-231 and BT474 cells. Relative intracellular expression of Ki67 RNA in T-47D cells ( A ), MDA-MB-231 cells ( B ) and BT474 cells ( C ) after treatment with 50µM citral (Cit), 50µM citrathal R (CR) and 50µM Cyclovertal (Cy). CTRL: DMSO-Control.

Article Snippet: BT474 cells (#300131) and T-47D cells (#300353) were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany), MDA-MB-231 cells (#92020424) from Sigma-Aldrich (Taufkirchen, Germany).

Techniques: Expressing

Journal: Breast Cancer : Targets and Therapy

Article Title: Anticarcinogenic Effects of Odorant Substances Citral, Citrathal R and Cyclovertal on Breast Cancer in vitro

doi: 10.2147/BCTT.S322619

Figure Lengend Snippet: Survivin in T-47D, MDA-MB-231 and BT474 cells. Relative intracellular expression of survivin RNA in T-47D cells ( A ), MDA-MB-231 cells ( B ) and BT474 cells ( C ) after treatment with 50µM citral (Cit), 50µM citrathal R (CR) and 50µM Cyclovertal (Cy). CTRL: DMSO-Control.

Article Snippet: BT474 cells (#300131) and T-47D cells (#300353) were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany), MDA-MB-231 cells (#92020424) from Sigma-Aldrich (Taufkirchen, Germany).

Techniques: Expressing

Journal: Breast Cancer : Targets and Therapy

Article Title: Anticarcinogenic Effects of Odorant Substances Citral, Citrathal R and Cyclovertal on Breast Cancer in vitro

doi: 10.2147/BCTT.S322619

Figure Lengend Snippet: Cyclin A2 in T47-D, MDA-MB-231 and BT474 cells. Relative intracellular expression of cyclin A2 RNA in T-47D cells ( A ), MDA-MB-231 cells ( B ) and BT474 cells ( C ) after treatment with 50µM citral (Cit), 50µM citrathal R (CR) and 50µM Cyclovertal (Cy). CTRL: DMSO-Control.

Article Snippet: BT474 cells (#300131) and T-47D cells (#300353) were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany), MDA-MB-231 cells (#92020424) from Sigma-Aldrich (Taufkirchen, Germany).

Techniques: Expressing

Journal: Breast Cancer : Targets and Therapy

Article Title: Anticarcinogenic Effects of Odorant Substances Citral, Citrathal R and Cyclovertal on Breast Cancer in vitro

doi: 10.2147/BCTT.S322619

Figure Lengend Snippet: Western blots. Western Blot showing control, capsazepine alone, citral, citral + capsazepine, citrathal R, citrathal R + capsazepine, cyclovertal and cyclovertal + capsazepine. ß-tubulin served as loading control in ( A ) T-47D, ( B ) BT474, ( C ) MDA-MB-231.

Article Snippet: BT474 cells (#300131) and T-47D cells (#300353) were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany), MDA-MB-231 cells (#92020424) from Sigma-Aldrich (Taufkirchen, Germany).

Techniques: Western Blot

Journal: Breast Cancer : Targets and Therapy

Article Title: Anticarcinogenic Effects of Odorant Substances Citral, Citrathal R and Cyclovertal on Breast Cancer in vitro

doi: 10.2147/BCTT.S322619

Figure Lengend Snippet: Immunocytochemistry. 1: T-47D PCNA. Immunocytochemical staining of T47D cells with PCNA antibody (dilution 1:7500, microscopic enlargement 400x). (a) control, (b) citral, (c) citrathal R, (d) cyclovertal. Compared to control reduced nuclear–cytoplasmic ratio and increased nuclear condensation. 2: T-47D Annexin V. Immunocytochemical staining of T47D cells with Annexin V antibody (dilution 1:250, microscopic enlargement 400x). (a) control, (b) citral, (c) citrathal R, (d) cyclovertal. Compared to control reduced nuclear–cytoplasmic ratio and increased nuclear condensation. Slightly increased staining of the outer cell membrane after citral treatment, more after citrathal R and cyclovertal, compared to the control. 3: BT474 PCNA. Immunocytochemical staining of BT474 cells with PCNA antibody (dilution 1:7500, microscopic enlargement 400x). (a) control, (b) citral, (c) citrathal R, (d) cyclovertal. Compared to control reduced nuclear–cytoplasmic ratio and increased nuclear condensation. 4: BT474 Annexin V. Immunocytochemical staining of BT474 cells with Annexin V antibody (dilution 1:250, microscopic enlargement 400x). (a) control, (b) citral, (c) citrathal R, (d) cyclovertal. Clearly increased staining of the outer cell membrane after citral, cylovertal and mostly after citrathal R treatment compared to the control. 5: MDA-MB-231 PCNA. Immunocytochemical staining of MDA-MB-231 cells with PCNA antibody (dilution 1:7500, microscopic enlargement 400x). (a) control, (b) citral, (c) citrathal R, (d) cyclovertal. Compared to control reduced condensation and increased fragmentation of the nucleus and loss of spindle-shape after treatment mostly with citral and citrathal R, but also cyclovertal. Less intensive staining of the nuclei after treatment with citral, citrathal R and cyclovertal compared to the control. 6: MDA-MB-231 Annexin V. Immunohistochemical staining of MDA-MB-231 cells with Annexin V antibody (dilution 1:250, microscopic enlargement 400x). (a) control, (b) citral, (c) citrathal R, (d) cyclovertal. Compared to control reduced condensation and increased fragmentation of the nucleus and loss of spindle-shape after treatment with citral, citrathal R and cyclovertal. Increased staining of the outer cell membrane after citral, citrathal R and mostly after cyclovertal treatment compared to the control.

Article Snippet: BT474 cells (#300131) and T-47D cells (#300353) were purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany), MDA-MB-231 cells (#92020424) from Sigma-Aldrich (Taufkirchen, Germany).

Techniques: Immunocytochemistry, Staining, Immunohistochemical staining

Journal: Nucleic acids research

Article Title: Spatially multiplexed RNA in situ hybridization to reveal tumor heterogeneity.

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: Figure 1. Local RNA in situ hybridization. (A) Schematic overview of a microfluidic implementation of chromogenic RNA in situ hybridization. The initial preparation of the tissue section including paraffin removal (1), heat (2) and protease (3) pretreatment, as well as fixation was performed globally. The primary probe was then hybridized locally to the RNA of interest (4). In this case the detection of HER2 is shown with internal negative (dapB) and positive (ActB) controls. Finally, the amplification of the signal using preamplifier and amplifier strands (5), the binding of enzyme (6), and the enzymatic reactions (7) were performed globally on the whole slide. (B) Fluorescence images of RNA-ISH experiments detecting the gene expression of HER2 in cell pellet sections of the breast cancer cell lines MCF7 (left), SKBR3 (center) and BT474 (right). Probes directed against bacterial dapB and ActB were used as negative and positive controls, respectively. Scale bar: 100 m.

Article Snippet: FFPE cell blocks and tissues FFPE cell blocks of the cell lines MCF7, BT474 and SKBR3 (AMS Biotechnology, Abingdon, UK) with HER2 expression levels of 1+, 2+ and 3+, respectively, were used as references.

Techniques: RNA In Situ Hybridization, Amplification, Binding Assay, Fluorescence, Gene Expression

Journal: Nucleic acids research

Article Title: Spatially multiplexed RNA in situ hybridization to reveal tumor heterogeneity.

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: Figure 2. Detection of HER2 expression status by RNA in situ hybridization using internal positive and negative controls. (A) Fluorescence images of the signals detected for HER2, dapB, and ActB in a mammary carcinoma section. Scale bar: 100 m. (B) Quantitative analysis of the fluorescence intensity of FISH signals detected for HER2, dapB, and ActB integrated per sectioned cell for FFPE sections of the breast cancer cell lines MCF7, SKBR3, and BT474 (see Figure 1) as well as the mammary carcinoma from (A). 100 ms excitation.

Article Snippet: FFPE cell blocks and tissues FFPE cell blocks of the cell lines MCF7, BT474 and SKBR3 (AMS Biotechnology, Abingdon, UK) with HER2 expression levels of 1+, 2+ and 3+, respectively, were used as references.

Techniques: Expressing, RNA In Situ Hybridization, Fluorescence

Journal: Nucleic acids research

Article Title: Spatially multiplexed RNA in situ hybridization to reveal tumor heterogeneity.

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: Figure 3. Single color multiplexed RNA-ISH for the detection of breast cancer biomarkers. (A) The primary probes for ER, PgR, and HER2 were delivered to spatially distinct regions of a mammary carcinoma section using a microfluidic chip (schematic in upper left corner). Fluorescence images of the signal detected for the breast cancer biomarkers ER, PgR and HER2 in a mammary carcinoma section. Scale bar: 100 m. (B) Fluorescence intensity of FISH signals detected for ER, PgR and HER2 integrated per sectioned cell for FFPE sections of the breast cancer cell lines MCF7, SKBR3 and BT474 (see Supplementary Figure S7) as well as the mammary carcinoma from (A). 500 ms excitation.

Article Snippet: FFPE cell blocks and tissues FFPE cell blocks of the cell lines MCF7, BT474 and SKBR3 (AMS Biotechnology, Abingdon, UK) with HER2 expression levels of 1+, 2+ and 3+, respectively, were used as references.

Techniques: Fluorescence