Journal:

Article Title: The SANT2 Domain of the Murine Tumor Cell DnaJ-like Protein 1 Human Homologue Interacts with ? 1 -Antichymotrypsin and Kinetically Interferes with Its Serpin Inhibitory Activity *

doi: 10.1074/jbc.M310903200

Figure Lengend Snippet: A, schematic representation of HTJ1 constructs used in this study. The position of amino acids in HTJ1, the size in kDa, and the interaction with ACT are indicated. B, immunoblot of total yeast protein extracts. Protein extracts from the AH109 yeast strain transformed with (lane 1), pGBKT7/pGBKT7/HTJ1242–554 HTJ1242–493 (lane 2), pGBKT7/HTJ1493–554 (lane 3), pGBKT7/HTJ1493–554:W497A (lane 4), pGBKT7/HTJ1493–554:W520A (lane 5), pGBKT7/HTJ1493–554:W497A, W520A (lane 6), pGBKT7/HTJ1242–411 (lane 7), pGBKT7/HTJ145–148 (lane 8), and pGBKT7 vector (lane 9) were separated on a 10% SDS-PAGE gel and detected with an anti-c-Myc monoclonal antibody.

Article Snippet: All components of the two-hybrid system, including cloning vectors pGBKT7 and pACT2, human liver cDNA library, S. cerevisiae strain AH109, anti-c-Myc monoclonal antibody, anti-HA tag polyclonal antibody, X-α-gal, dropout supplements, and cobalt-chelating resin Talon were obtained from Clontech.

Techniques: Construct, Western Blot, Transformation Assay, Plasmid Preparation, SDS Page

Journal: OncoTargets and Therapy

Article Title:

Piwi-Interacting RNA1037 Enhances Chemoresistance and Motility in Human Oral Squamous Cell Carcinoma Cells

doi: 10.2147/ott.s233322

Figure Lengend Snippet: Figure 2 Targeting the upregulated expression of piR-1037 induced by CDDP enhanced the sensitivity of OSCC cells to CDDP. (A) Confirmation of the suppressive effect of piRNA-1037 antisense oligonucleotides (piR-1037 complementary DNA oligonucleotides) (500 nM) on the expression of piRNA-1037 in SCC4 and SCC9 cells receiving the indicated treatments. (B) Western blot analysis and quantification of the effect of piR-1037 antisense oligonucleotides on the expression of the chemoresistance biomarkers MDR1 and α-enolase in SCC4 and SCC9 cells treated with CDDP (10 μM). (C) Decreased cell viability and increased apoptosis (TUNEL assay) (D) in OSCC cells transfected with piR-1037 antisense oligonucleotides and treated with CDDP compared with CDDP-treated cells transfected with a scrambled control. Cell apoptosis was detected using the TUNEL-based TiterTACS™Colorimetric Apoptosis Detection Kit (Trevigen, USA). (E) Activity of caspase-3, caspase-8 and caspase-1 (F). The activity of caspases measured using Caspase-3 and Caspase-1 assay kits (Abcam, USA) and a Caspase-8 colorimetric kit (Sigma Aldrich, USA). The data represent the mean ±SD from at least three independent replicates. Statistical analysis was performed using one-way ANOVA analysis or unpaired student’s t-test: *p <0.05; **p < 0.01.

Article Snippet: The membrane was then incubated with 5% nonfat milk in TBST (10 mM Tris, pH 8.0, 150 mM NaCl, 0.5% Tween 20) for 1 h. The membrane was washed once with TBST, followed by incubation with primary antibodies against MDR1 (Novus Biologicals, USA) (1:1000), α-enolase (Boster, China) (1:1000), XIAP (Abcam, USA) (1 μg/mL), cleaved caspase-3 (R&D systems, USA) (0,5 μg/mL), E-Cadherin (1:500), N-Cadherin (1:1000) (Abcam, USA), and β-actin (1:10,000) (Sigma Aldrich, USA).

Techniques: Expressing, Western Blot, TUNEL Assay, Transfection, Control, Activity Assay