Journal: Nucleic Acids Research

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

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: 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 μm.

Article Snippet: 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, Expressing

Journal: Nucleic Acids Research

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

doi: 10.1093/nar/gkz1151

Figure Lengend Snippet: 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 μm. ( 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 ) as well as the mammary carcinoma from (A). 100 ms excitation.

Article Snippet: 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: 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 μm. ( 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 ) as well as the mammary carcinoma from (A). 500 ms excitation.

Article Snippet: 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

Journal: Membranes

Article Title: Liposomes Loaded with the Proteasome Inhibitor Z-Leucinyl-Leucinyl-Norleucinal Are Effective in Inducing Apoptosis in Colorectal Cancer Cell Lines

doi: 10.3390/membranes10050091

Figure Lengend Snippet: LLNle causes a concentration-dependent inhibition of survival in Caco-2, GP2d and LoVo cell lines. An MTT assay was performed after 72 h of treatment of the indicated cell culture, n = 4 replicas were performed and average values and SD are shown for each concentration indicated on the x -axis. An interpolation curve of survival values is shown for the GP2d cell line.

Article Snippet: CRC cell lines Caco-2 and LoVo were obtained from Banca Biologica and Cell Factory in IRCCS Ospedale Policlinico San Martino, belonging to the European Culture Collection’s Organization.

Techniques: Concentration Assay, Inhibition, MTT Assay, Cell Culture

Journal: Membranes

Article Title: Liposomes Loaded with the Proteasome Inhibitor Z-Leucinyl-Leucinyl-Norleucinal Are Effective in Inducing Apoptosis in Colorectal Cancer Cell Lines

doi: 10.3390/membranes10050091

Figure Lengend Snippet: LLNLe induces apoptosis in Caco-2, GP2d and LoVo cell lines. Cells were treated for 72 h with LLNle (1.7, 1.4 and 0.6 µM LLNle for Caco-2, GP2d and LoVo, respectively) or with DMSO used as a control. Cells were labeled with both the DNA-specific and cell-impermeant sytox-blue, and annexin-V APC and fluorescence emissions were analyzed by FCM. Early apoptotic (annexin-V APC + and sytox-blue − ) cells are indicated by red arrows while necrotic cells (annexin-V APC − and sytox-blue + ) are indicated by a black arrow.

Article Snippet: CRC cell lines Caco-2 and LoVo were obtained from Banca Biologica and Cell Factory in IRCCS Ospedale Policlinico San Martino, belonging to the European Culture Collection’s Organization.

Techniques: Control, Labeling, Fluorescence

Journal: Membranes

Article Title: Liposomes Loaded with the Proteasome Inhibitor Z-Leucinyl-Leucinyl-Norleucinal Are Effective in Inducing Apoptosis in Colorectal Cancer Cell Lines

doi: 10.3390/membranes10050091

Figure Lengend Snippet: LLNle-liposome inhibits CRC cell line survival. MTT analysis of Caco-2, GP2d, and LoVo cell line treated with LLNle, DMSO (used as a control for LLNle), LLNle-liposomes, Cxm-liposomes-LLNle, empty liposomes, and PBS (control for liposomes) for 72 h. LLNle-liposomes were diluted to achieve an LLNle final concentration in the culture medium equivalent to 2.5, 2.0 and 0.8 µM of free LLNle for Caco-2, GP2d and LoVo, respectively. All measurements here reported are presented as mean ± standard deviations (s.d.), n = 4. Details of the results of the statistical analysis are provided in .

Article Snippet: CRC cell lines Caco-2 and LoVo were obtained from Banca Biologica and Cell Factory in IRCCS Ospedale Policlinico San Martino, belonging to the European Culture Collection’s Organization.

Techniques: Control, Liposomes, Concentration Assay

Journal: Membranes

Article Title: Liposomes Loaded with the Proteasome Inhibitor Z-Leucinyl-Leucinyl-Norleucinal Are Effective in Inducing Apoptosis in Colorectal Cancer Cell Lines

doi: 10.3390/membranes10050091

Figure Lengend Snippet: LLNle-liposome inhibits proteasomal degradation of poly-ubiquitinated protein and induces apoptosis in CRC cell lines. Immunoblot analysis of whole-cell lysates of Caco-2, GP2d, and LoVo cell lines treated with LLNle, DMSO (used as control for LLNle), LLNle-liposomes, and PBS (control for LLNle-liposomes) for 72 h. LLNle-liposomes were diluted to achieve an LLNle final concentration in the culture medium equivalent to 1.7, 1.4 and 0.6 µM LLNle of free LLNle for Caco-2, GP2d and LoVo, respectively. All three cell lines display expression of ERBB1, the EGF receptor. A high molecular weight smear signal, corresponding to poly-ubiquitinated proteins, and the generation of the 89 kDa isoform of the protein PARP was detected by specific Ab only in LLNle-liposomes and LLNle treated samples. Tubulin was used as a loading control.

Article Snippet: CRC cell lines Caco-2 and LoVo were obtained from Banca Biologica and Cell Factory in IRCCS Ospedale Policlinico San Martino, belonging to the European Culture Collection’s Organization.

Techniques: Western Blot, Control, Liposomes, Concentration Assay, Expressing, High Molecular Weight

Journal: Diagnostic Pathology

Article Title: Overexpression of c- erb B2 is a negative prognostic factor in anaplastic astrocytomas

doi: 10.1186/1746-1596-5-18

Figure Lengend Snippet: erb B antibodies used

Article Snippet: c- erb B-2 , Novocastra , Monoclonal , CB11 , c- erb B2 protein (internal domain) , SKBR-3 breast cancer cell line , 1:40 , 60 min. at room temperature.

Techniques: Positive Control, Incubation