paraffin section of tumor tissues  (Servicebio Inc)

Journal: Frontiers in Oncology

Article Title: Salubrinal Exposes Anticancer Properties in Inflammatory Breast Cancer Cells by Manipulating the Endoplasmic Reticulum Stress Pathway

doi: 10.3389/fonc.2021.654940

Figure Lengend Snippet: BiP/GRP78 is expressed abundantly in human IBC tissue sections. (A) Hematoxylin and Eosin (H & E) staining of healthy (left) (Aa) and IBC tissue (right) (Ab) is shown at 10X and 40X magnification. Healthy (Ac, Ad) and IBC tissue sections (Ae, Af) were stained using an antibody against BiP/GRP78. Human IBC tissue sections (Ag, Ah) were stained using an anti-IgG antibody. (Aa–Ac, Ae, Ag) are 10X, whereas (Ad, Af, Ah) represent 40X magnification. (B) BiP/GRP78 immunostaining (red) and healthy and IBC tissue are shown at 10X magnification. Tissue sections were developed with Alexa-594 coupled secondary antibody (red). Nuclei were visualized using DAPI as the counterstain (blue).

Article Snippet: Total RNA was isolated using TRIzol Reagent (#15596026, Life Technologies Corporation, Grand Island, NY) from IBC tissue samples (Biochain, breast tumor tissue array # T22350862-2) and treated with DNase I (#18068015, Life Technologies Corporation) at 37°C for 30 min for DNA removal.

Techniques: Staining, Immunostaining

Journal: Frontiers in Oncology

Article Title: Salubrinal Exposes Anticancer Properties in Inflammatory Breast Cancer Cells by Manipulating the Endoplasmic Reticulum Stress Pathway

doi: 10.3389/fonc.2021.654940

Figure Lengend Snippet: ER stress genes are abundantly expressed in IBC tissue sections and IBC cell lines. (A) RNA isolated from healthy and IBC tumor tissue was prepared, converted to cDNA, and gene expression was quantified by real-time RT-PCR using specific primers for ER stress markers as indicated. Each data point represents the average gene expression from six IBC and six healthy control samples. Each point represents the average ± the standard deviation of three experiments. (***) p<0.005, (****) p<0.001 indicates a statistically significant difference compared with healthy tissue. Each reaction was done in triplicate. (B) RNA isolated from HMEC, SUM149PT, and SUM190PT cells was converted to cDNA, and gene expression was quantified by real-time RT-PCR using specific primers for ATF4, CHOP, GADD34, GRP78, IRE1α, and XBP-1. Each point represents the average ± the standard deviation of three experiments. (**) p<0.01, (***) p<0.005, (****) p<0.001 indicate a statistically significant difference compared with HMEC cells. Each reaction was done in triplicate. (C) ER stress genes proteins are abundantly expressed in IBC cell lines. Lysates prepared from HMEC, SUM149PT, and SUM190PT cells, were tested for protein levels of PERK, IRE1α, calnexin, ERO1α, and PDI. Blots were reprobed with anti-β-actin antibody as a loading control for normalization. Fold expression of each protein was calculated by considering the expression of the protein in HMEC as 1. (D) Immunostaining of HMEC and SUM149PT cells seeded in eight-well chamber slides. Cells were fixed, permeabilized, and then stained with primary monoclonal antibodies against ER stress markers, including calnexin, ERO1α, and IRE1α. Cells were developed with Alexa-488 coupled secondary antibody (green). Nuclei were visualized using DAPI as the counterstain (blue).

Article Snippet: Total RNA was isolated using TRIzol Reagent (#15596026, Life Technologies Corporation, Grand Island, NY) from IBC tissue samples (Biochain, breast tumor tissue array # T22350862-2) and treated with DNase I (#18068015, Life Technologies Corporation) at 37°C for 30 min for DNA removal.

Techniques: Isolation, Expressing, Quantitative RT-PCR, Standard Deviation, Immunostaining, Staining

Journal: Cancer Research and Treatment : Official Journal of Korean Cancer Association

Article Title: Activation of β 2 -Adrenergic Receptor Promotes Growth and Angiogenesis in Breast Cancer by Down-regulating PPARγ

doi: 10.4143/crt.2019.510

Figure Lengend Snippet: β 2 receptor activation and peroxisome proliferator-activated receptor γ (PPARγ) inhibition is involved in chronic stress-induced tumor angiogenesis in vivo . (A) Representative tumor images from mice in control (n=4), ICI118551 (n=4) and GW9662 group (n=5). (B) Tumor growth curves. (C) Average tumor weight was measured on day 28. * p < 0.05 vs. control, # p < 0.05 vs. chronic stress. (D, E) PPARγ, vascular endothelial growth factor (VEGF), and fibroblast growth factor 2 (FGF2) mRNA expression in tumor tissues by reverse transcription polymerase chain reaction (D) and quantitative real-time polymerase chain reaction (E). * p < 0.05. (F) PPARγ, VEGF, and FGF2 protein expression in tumor tissues by western blot. GAPDH, glyceraldehyde 3-phosphate dehydrogenase. (G) Representative images (left panel) and bar graph (right panel) of PPARγ expression detected in tumor tissue paraffin sections by immunohistochemistry staining. * p < 0.05. (H) Images of tumor section CD31 immunohistochemistry staining (left panel) and microvascular density was measured (right panel). * p < 0.05 vs. control, # p < 0.05 vs. chronic stress.

Article Snippet: For immunohistochemistry, tumor tissue paraffin sections from tumor-bearing mice were stained with anti-CD31 (1:100, sc-71873, Santa Cruz Biotechnology) or anti-PPARγ primary antibodies (1:100).

Techniques: Activation Assay, Inhibition, In Vivo, Control, Expressing, Reverse Transcription, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Immunohistochemistry, Staining