Journal: BMC Musculoskeletal Disorders

Article Title: Type-1 Collagen differentially alters β-catenin accumulation in primary Dupuytren's Disease cord and adjacent palmar fascia cells

doi: 10.1186/1471-2474-10-72

Figure Lengend Snippet: Type 1 collagen culture effects on β-catenin accumulation in DD and PF-cells . PF cells (A) and DD cells (B) were cultured on Type 1 collagen coated slides (1.9 mg/ml) for 72 hours. Cells were fixed and stained with Oregon green phalloidin (green) and DAPI (blue) to label filamentous actin and the nucleus, respectively. β-catenin was detected using primary beta-catenin and TRITC-conjugated secondary antibodies (orange). As shown, PF cells consistently displayed readily detectable peri-nuclear staining as well as diffuse staining throughout the cytoplasm after three days on collagen culture. In contrast, DD cells displayed distinct β-catenin staining at the periphery of some cells adjacent to the cell membrane, with little or no β-catenin evident in the cytoplasm after three days on collagen culture.

Article Snippet: Cells were stained for DNA (DAPI) actin stress fibers (Alexa 488 phallodin, Molecular Probes, Eugene OR), and β-catenin (TRITC-conjugated anti-β-catenin, BD Biosciences, Mississauga ON).

Techniques: Cell Culture, Staining, Membrane

Journal: Cancer Management and Research

Article Title: Increased expression of TMED2 is an unfavorable prognostic factor in patients with breast cancer

doi: 10.2147/CMAR.S192949

Figure Lengend Snippet: The expression level of TMED2 in breast cancer and normal breast cancer. ( A ) The expression of TMED2 was elevated in breast cancer compared to normal breast tissues. Data derived from UALCAN database. ( B–C ) Immunohistochemical analysis of TMED2 expression in breast cancer and normal mammary tissue and overview of TMED2 protein levels in breast tumor tissues and normal breast tissues. Data derived from the Human Protein Atlas database. ( D ) The TMED2 protein expression of breast cancer tissue (a–c), breast normal tissue (d–f), SCs and NSCs were examined by Western Blot and β-actin was used as an internal reference. Data were from 3 independent experiments. ( E ) The DNA methylation level of breast cancer and normal breast tissue, the Beta value indicates level of DNA methylation ranging from 0 (unmethylated) to 1 (fully methylated). Different beta value cut-off has been considered to indicate hyper-methylation. Data derived from UALCAN database. ( F ) The DNA copy number of breast cancer and normal breast tissue and data derived from oncomine database and analyzed with GraphPad.

Article Snippet: The membranes were blocked with 5% low-fat milk in TBST for 60 min and then incubated with the following primary antibodies overnight (4 °C): TMED2 (241213, Abcam,1:2000), β-actin (6276, Abcam,1:5000); The corresponding secondary antibodies (1:5000) were added and bands were visualized by enhanced chemiluminescence (PerkinElmer, MA, USA).

Techniques: Expressing, Derivative Assay, Immunohistochemical staining, Western Blot, DNA Methylation Assay, Methylation

Journal: Cell Reports Medicine

Article Title: Targeting pancreatic cancer glutamine dependency confers vulnerability to GPX4-dependent ferroptosis

doi: 10.1016/j.xcrm.2025.101928

Figure Lengend Snippet:

Article Snippet: HRP-conjugated Beta Actin , Proteintech , Cat# HRP-60008; RRID: AB_2819183.

Techniques: Virus, CRISPR, Recombinant, Protease Inhibitor, Lysis, Ligation, Reverse Transcription, SYBR Green Assay, Glo Assay, Sample Prep, DNA Extraction, Enzyme-linked Immunosorbent Assay, Multiple Displacement Amplification, Flow Cytometry, Software

Journal: Frontiers in Microbiology

Article Title: KAT2A Promotes Hepatitis B Virus Transcription and Replication Through Epigenetic Regulation of cccDNA Minichromosome

doi: 10.3389/fmicb.2021.795388

Figure Lengend Snippet: Identification of KAT2A as a host factor promoting cccDNA transcription. (A) The reported succinyltransferases and desuccinylases were summarised in the table. (B) After 4 days of siRNA treatment, total RNA was extracted by TRNzol Universal reagent. The silencing efficiency of related genes in Huh-7 cells was analysed by real-time PCR with specific primers. β-actin was used as the internal control. (C) The indicated siRNA (50 pmol) and monomeric linearised HBV DNA (1 μg) were co-transfected into huh-7 cells by using Lipofectamine™ 3000 Transfection Reagent. The supernatant was collected and HBeAg and HBsAg levels were detected by ELISA. * P < 0.05.

Article Snippet: Mouse anti-HBcAg monoclonal antibody (sc-23945), mouse anti-GAPDH monoclonal antibody (sc-47724) and rabbit anti-β-actin monoclonal antibody (sc-47778) were obtained from Santa Cruz Biotechnology (United States).

Techniques: Real-time Polymerase Chain Reaction, Transfection, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Microbiology

Article Title: KAT2A Promotes Hepatitis B Virus Transcription and Replication Through Epigenetic Regulation of cccDNA Minichromosome

doi: 10.3389/fmicb.2021.795388

Figure Lengend Snippet: KAT2A knockdown suppresses cccDNA transcription in the HBV infection cell model. HepG2-NTCP cells were infected HBV particles at 1-day post shRNA transduction. At 4 days after infection. (A) The expression of KAT2A was detected by western blot. (B) Northern blotting and real-time PCR were used to analyse the effect of KAT2A knockdown on HBV RNAs. (C) HBV cccDNA was extracted by a modified Hirt DNA extraction protocol and then Southern blotting and Taq-man probe qPCR were used to detect the HBV cccDNA level. The mitochondrial gene Cox1 was hybridised as the loading control for HBV cccDNA. (D) The level of HBV 3.5-kb RNA, total HBV RNAs, and cccDNA were used to calculate the ratio of HBV 3.5-kb RNA to cccDNA and total HBV RNAs to cccDNA. (E) HBV core DNA was quantified by Southern blotting and absolute quantification PCR. The level of β-actin was used as a loading control for HBV core DNA. (F) HBV core protein (HBc) was produced by HBV infected HepG2-NTCP cells. Intracellular HBc was visualised by immunofluorescence staining. The scale bar is 100 μm. (G) Western blot was used to detect the level of HBsAg in cells (left). Cell culture supernatant was collected for HBsAg and HBeAg analysis via ELISA (right). Cox1, cyclooxygenase 1; rRNA, ribosomal RNA. * P < 0.05.

Article Snippet: Mouse anti-HBcAg monoclonal antibody (sc-23945), mouse anti-GAPDH monoclonal antibody (sc-47724) and rabbit anti-β-actin monoclonal antibody (sc-47778) were obtained from Santa Cruz Biotechnology (United States).

Techniques: Infection, shRNA, Transduction, Expressing, Western Blot, Northern Blot, Real-time Polymerase Chain Reaction, Modification, DNA Extraction, Southern Blot, Produced, Immunofluorescence, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Microbiology

Article Title: KAT2A Promotes Hepatitis B Virus Transcription and Replication Through Epigenetic Regulation of cccDNA Minichromosome

doi: 10.3389/fmicb.2021.795388

Figure Lengend Snippet: KAT2A overexpression promotes HBV transcription in the HBV infection cell model. HepG2-NTCP cells were infected HBV particles at 1-day post lentivirus expressing KAT2A plasmids transduction. At 4 days after infection. (A) The expression of KAT2A was detected by western blot. (B) The effect of KAT2A overexpression on HBV RNAs was analysed by Northern blotting and real-time PCR. (C) Taq-man probe qPCR was used to detect the HBV cccDNA level. (D) The level of HBV 3.5-kb RNA, total HBV RNAs, and cccDNA were used to calculate the ratio of HBV 3.5-kb RNA to cccDNA and total HBV RNAs to cccDNA. (E) HBV core DNA was quantified by Southern blotting and absolute quantification PCR. The level of β-actin was used as a loading control for HBV core DNA. (F) HBc was produced by HBV-infected HepG2-NTCP cells. Intracellular HBc was visualised by immunofluorescence staining. The scale bar is 100 μm. (G) Cell culture supernatant was collected for HBsAg and HBeAg analysis via ELISA. * P < 0.05.

Article Snippet: Mouse anti-HBcAg monoclonal antibody (sc-23945), mouse anti-GAPDH monoclonal antibody (sc-47724) and rabbit anti-β-actin monoclonal antibody (sc-47778) were obtained from Santa Cruz Biotechnology (United States).

Techniques: Over Expression, Infection, Expressing, Transduction, Western Blot, Northern Blot, Real-time Polymerase Chain Reaction, Southern Blot, Produced, Immunofluorescence, Staining, Cell Culture, Enzyme-linked Immunosorbent Assay