Journal:

Article Title: Changes in the Expression of Genes Associated with Intraneuronal Amyloid Beta and Tau in Alzheimer's Disease

doi: 10.3233/JAD-2010-1216

Figure Lengend Snippet: Hippocampal cells coexpressing Aβ and TrkB are present in AD and control cases. Representative DIF of Aβ/TrkB showing high levels of Aβ (green) in AD (AD5) compared to control (C5); higher levels of TrkB (red) in control compared to AD; and similar numbers of Aβ/TrkB (yellow cells in merge) in controls compared to AD (×200). Arrows point to examples of coexpressing cells. The bottom two panels are the negative controls using nonspecific mouse or rabbit IgG as primary antibodies and the appropriate FITC-avidin or Texas Red DCS. No immunostaining was detected.

Article Snippet: After incubation with antibodies to either Aβ or P-tau, antibody binding was detected with biotinylated anti-rabbit or mouse IgG (1/200 dilution; 20 min at room temperature; Vector Laboratories, Burlingame, CA), a 1:300 dilution of FITC-avidin DCS (Vector Laboratories) for 5 min, followed by avidin/biotin blocking for 15 min, and then immunostaining with BDNF, TrkB, or DYN and detection with biotinylated anti-rabbit or mouse IgG and Texas Red DCS for 5 min. Psuedocolor images were captured using a Nikon fluorescence microscope with FITC and Texas Red filters using IP Lab software (Fairfax, VA) and merged using Adobe Photoshop (San Jose, CA).

Techniques: Avidin-Biotin Assay, Immunostaining

Journal: Molecular medicine reports

Article Title: Knockdown of biglycan expression by RNA interference inhibits the proliferation and invasion of, and induces apoptosis in, the HCT116 colon cancer cell line.

doi: 10.3892/mmr.2015.4383

Figure Lengend Snippet: Figure 1. Construction of the stable HCT116 cell line with biglycan down regulation. The (A) mRNA and (B) protein expression levels of biglycan in the shRNA‑biglycan/control‑transfected or non‑transfected cells was detected using reverse transcription‑quantitative polymerase chain reaction and western blotting, respectively. (C) The relative protein expression of big lycan was normalized against β‑actin. Each experiment was repeated three times and the data are expressed as the mean ± standard deviation (*P<0.05 or **P<0.01, compared with the shRNA‑control group). sh, short hairpin.

Article Snippet: The membrane was subsequently blocked with 5% non-fat dry milk at room temperature for 1 h, followed by incubation with the following diluted primary antibodies: Rabbit polyclonal anti-biglycan antibody (cat no. sc-33788; 1:100; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), rabbit polyclonal anti-caspase-3 antibody (cat no. wl01992a; 1:1,000), rabbit polyclonal anti-p27 antibody (cat no. wl01769; 1:1,000), rabbit polyclonal anti-cyclin A antibody (cat no. wl01753; 1:200), rabbit polyclonal anti-p21 antibody (cat no. wl0362; 1:100), rabbit polyclonal anti-cyclin D1 antibody (cat no. wl01435a; 1:100) (all from Wanleibio, Shenyang, China), rabbit polyclonal anti-p38 antibody (cat no. sc-7149; 1:100) and anti-phosphorylated-p38 antibody (cat no. sc-101758; 1:100) (both from Abcam, Cambridge, MA, USA) at 4 ̊C overnight.

Techniques: Expressing, Polymerase Chain Reaction, Western Blot, Standard Deviation

Journal: Molecular medicine reports

Article Title: Knockdown of biglycan expression by RNA interference inhibits the proliferation and invasion of, and induces apoptosis in, the HCT116 colon cancer cell line.

doi: 10.3892/mmr.2015.4383

Figure Lengend Snippet: Figure 2. Downregulation of biglycan inhibits the proliferation of colon cancer cells and causes cell cycle arrest. (A) The effect of the downregulation of biglycan on HCT116 cell proliferation was measured using a cell counting kit‑8 assay, with each group containing six technical replicates. (B and C) The effect of the downregulation of biglycan on the HCT116 cell cycle distribution was analyzed by flow cytometry. (D) The protein expression levels of cyclin A, cyclin D1, p21 and p27 were detected by western blotting, and the results obtained from a representative experiment are shown. (E) The relative protein expres sion of these proteins were normalized against β‑actin. The data are expressed as the mean ± standard deviation (**P<0.01, compared with the shRNA‑control group). OD, optical density; sh, short hairpin.

Article Snippet: The membrane was subsequently blocked with 5% non-fat dry milk at room temperature for 1 h, followed by incubation with the following diluted primary antibodies: Rabbit polyclonal anti-biglycan antibody (cat no. sc-33788; 1:100; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), rabbit polyclonal anti-caspase-3 antibody (cat no. wl01992a; 1:1,000), rabbit polyclonal anti-p27 antibody (cat no. wl01769; 1:1,000), rabbit polyclonal anti-cyclin A antibody (cat no. wl01753; 1:200), rabbit polyclonal anti-p21 antibody (cat no. wl0362; 1:100), rabbit polyclonal anti-cyclin D1 antibody (cat no. wl01435a; 1:100) (all from Wanleibio, Shenyang, China), rabbit polyclonal anti-p38 antibody (cat no. sc-7149; 1:100) and anti-phosphorylated-p38 antibody (cat no. sc-101758; 1:100) (both from Abcam, Cambridge, MA, USA) at 4 ̊C overnight.

Techniques: CCK-8 Assay, Flow Cytometry, Expressing, Western Blot, Standard Deviation

Journal: Molecular medicine reports

Article Title: Knockdown of biglycan expression by RNA interference inhibits the proliferation and invasion of, and induces apoptosis in, the HCT116 colon cancer cell line.

doi: 10.3892/mmr.2015.4383

Figure Lengend Snippet: Figure 3. Downregulation of biglycan suppresses the migratory and invasive properties of colon cancer cells (magnification, x200). (A) The motility of shRNA‑biglycan/control or non‑transfected cells was determined using a scratch wound assay over a 24 h time period. (B) The migration rate was determined by the distance traveled by the cells to the front of the denuded area. (C) The invasion ability of cells in each group was measured using a Transwell assay. (D) The number of invasive cells were counted under an inverted microscope and cell numbers were plotted. The results obtained from a representative experi ment are shown. The data are expressed as the mean ± standard deviation (**P<0.01, compared with the shRNA‑control group).. sh, short hairpin.

Article Snippet: The membrane was subsequently blocked with 5% non-fat dry milk at room temperature for 1 h, followed by incubation with the following diluted primary antibodies: Rabbit polyclonal anti-biglycan antibody (cat no. sc-33788; 1:100; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), rabbit polyclonal anti-caspase-3 antibody (cat no. wl01992a; 1:1,000), rabbit polyclonal anti-p27 antibody (cat no. wl01769; 1:1,000), rabbit polyclonal anti-cyclin A antibody (cat no. wl01753; 1:200), rabbit polyclonal anti-p21 antibody (cat no. wl0362; 1:100), rabbit polyclonal anti-cyclin D1 antibody (cat no. wl01435a; 1:100) (all from Wanleibio, Shenyang, China), rabbit polyclonal anti-p38 antibody (cat no. sc-7149; 1:100) and anti-phosphorylated-p38 antibody (cat no. sc-101758; 1:100) (both from Abcam, Cambridge, MA, USA) at 4 ̊C overnight.

Techniques: Control, Scratch Wound Assay Assay, Migration, Transwell Assay, Inverted Microscopy, Standard Deviation

Journal: Molecular medicine reports

Article Title: Knockdown of biglycan expression by RNA interference inhibits the proliferation and invasion of, and induces apoptosis in, the HCT116 colon cancer cell line.

doi: 10.3892/mmr.2015.4383

Figure Lengend Snippet: Figure 4. Downregulation of biglycan activates the p38 signaling pathway and induces apoptosis in colon cancer cells. (A) The effect of biglycan downregula tion on the apoptosis of the HCT116 cells was detected using flow cytometry, and results are shown from a representative experiment. (B) The proportion of apoptotic cells in each group was determined. (C) The protein expression levels of caspase‑3 and p‑p38 were detected by western blotting, and the results obtained from a representative experiment are shown for each group. (D) The relative expression of these proteins was normalized against β‑actin. The data are expressed as the mean ± standard deviation (**P<0.01, compared with the shRNA‑control group; ##P<0.01, compared with the shRNA‑biglycan group). p‑, phosphorylated; sh, short hairpin.

Article Snippet: The membrane was subsequently blocked with 5% non-fat dry milk at room temperature for 1 h, followed by incubation with the following diluted primary antibodies: Rabbit polyclonal anti-biglycan antibody (cat no. sc-33788; 1:100; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), rabbit polyclonal anti-caspase-3 antibody (cat no. wl01992a; 1:1,000), rabbit polyclonal anti-p27 antibody (cat no. wl01769; 1:1,000), rabbit polyclonal anti-cyclin A antibody (cat no. wl01753; 1:200), rabbit polyclonal anti-p21 antibody (cat no. wl0362; 1:100), rabbit polyclonal anti-cyclin D1 antibody (cat no. wl01435a; 1:100) (all from Wanleibio, Shenyang, China), rabbit polyclonal anti-p38 antibody (cat no. sc-7149; 1:100) and anti-phosphorylated-p38 antibody (cat no. sc-101758; 1:100) (both from Abcam, Cambridge, MA, USA) at 4 ̊C overnight.

Techniques: Flow Cytometry, Expressing, Western Blot, Standard Deviation