Journal: International journal of molecular sciences

Article Title: Apoptotic Caspases-3 and -7 Cleave Extracellular Domains of Membrane-Bound Proteins from MDA-MB-231 Breast Cancer Cells.

doi: 10.3390/ijms26083466

Figure Lengend Snippet: Figure 5. Immunological detection of caspase-mediated cleavages of extracellular proteins in normal (pH 7.4) and acidic (pH 6.0) environments. For the detection of extracellular cleavages, MDA-MB-231 cells were used. Red arrows indicate bands representing cleaved fragments, while black arrows indicate full-length proteins present in total cell lysates. (A) Detected caspase-3 or -7 extracellular cleavages of the selected targets (NRP-1, CD44, CSPG4) in an acidic (pH 6.0) environment in either DPBS or MES buffer. (B) Detection of caspase extracellular cleavages in a normal (pH 7.4) environment using either DPBS or HEPES buffer. NRP-1, neuropilin-1; CD44, CD44 antigen; CSPG4, chondroitin sulfate proteoglycan 4.

Article Snippet: Primary antibodies used for detection were polyclonal rabbit antibodies against cleaved caspase-3 (#9661, Cell Signaling, Danvers, MA, USA), polyclonal rabbit antibodies against cleaved caspase-7 (#9491, Cell Signaling, Danvers, MA, USA), polyclonal rabbit antibodies against β-actin (A2066, Sigma Aldrich, St. Louis, MO, USA), polyclonal sheep antibodies against human neuropilin-1 (#AF3870, R&D Systems, Minneapolis, MN, USA), monoclonal mouse antibodies against human CD44 (#3570, Cell Signaling, Danvers, MA, USA) and rabbit monoclonal antibodies against human CSPG4 (#43916, Cell Signaling, Danvers, MA, USA).

Techniques:

Journal: Aging (Albany NY)

Article Title: Cysteinyl leukotriene receptor 1 modulates retinal immune cells, vascularity and proteolytic activity in aged mice

doi: 10.18632/aging.206193

Figure Lengend Snippet: Pericyte count per capillary mm in the retinas of young untreated, vehicle-treated and MTK-treated old mice. The pericytes were visualized by NG2 labeling, and the capillaries were visualized by ColIV labeling. Capillary length was measured using ImageJ. ( A ) Pericyte count per capillary mm labeling in superficial retinal layers of young untreated, vehicle-treated and MTK-treated old mice and ( B ) representative NG2 (green) and ColIV (red) labeling in superficial retinal layers of young untreated mice. ( C ) Pericyte count per capillary mm in the deep retinal layers of young untreated, vehicle-treated and MTK-treated old mice and ( D ) representative NG2 and ColIV labeling in the deep retinal layers of young untreated mice. Scale bar = 100 µm. The data are represented as bar graphs and scatter plots ± SDs, n = 3–11. Two-way ANOVA (main factors: group and sex) followed by a Dunnett multiple comparison test.

Article Snippet: Afterward, the tissues were washed 3 times with TBS for 20 minutes and incubated in binding buffer containing specific antibodies against GFAP (1:500, GP52, Progen, Germany), Iba1 (1:300, 019-19741, FUJIFILM Wako Pure Chemical Corporation, Japan), NG2 (1:300, 481 005, Synaptic Systems, Germany), ColIV (1:300, AB769, Merck Millipore, MA, USA), Brn3a (1:200, sc-31984, Santa Cruz Biotechnology, TX, USA), Sqstm1 (1:100, #5114, Cell Signaling Technology, UK), cathepsin D (1:100, ab75852, Abcam, UK), Lamp1 (1:50, 18992, Santa Cruz), Lamp2a (1:200, ab18528, Abcam) and γH2AX (1:200, 9718, Cell Signaling) for 3 days at 4°C.

Techniques: Labeling, Comparison