Journal:

Article Title: Expression of RASSF1A, an epigenetically silenced tumor suppressor, overcomes resistance to apoptosis induction by interferons

doi: 10.1158/0008-5472.CAN-05-2303

Figure Lengend Snippet: A): Map of lentiviral RASSF1A construct. B): ACHN cells were transduced with RASSF1A carrying lentivirus or empty virus. 72h later selective antibiotic was added and the population of clones was grown in selection media for 14 days before assessing apoptotic response to IFN-β (50 U/ml over 5 days) by TUNEL assay. After confirmation of stable expression of RASSF1A by immunoblotting, clonal selection was performed to identify clones that expressed similar amounts of RASSF1A protein as achieved by DNMT1 AS (40 nM) over 8 days. Treatment with DNMT1 mismatch control oligonucleotide (MM) and empty lentivirus transduced cells served as negative controls. In clones expressing RASSF1A at levels comparable to DNA demethylating treatment, IFN-β (50 U/ml over 5 days) resulted in 20–40% apoptotic cells while stable transduction with empty lentivirus (kept in selective antibiotic for equal amounts of time) did not. C): Concurrent treatment of ACHN RASSF1A clone 1.3 with 2 μg/ml TRAIL neutralizing antibody (TRAIL-N AB) and IFN-β (50 U/ml) over 5 days inhibited IFN-induced apoptosis compared to cotreatment with 2 μg/ml control rabbit immunoglobulin (CTRL AB). D): RASSF1A expression markedly increased sensitivity to apoptosis induction by Apo2L/TRAIL in ACHN cells. Normal kidney epithelial (NKE) cells, even after pretreatment with 5-AZA-dC (200 nM) over 4 days, which alone resulted in moderate apoptosis induction, remained resistant to the apoptosis inducing effects of Apo2L/TRAIL. TUNEL graphs display the means and standard deviations (error bars) of independent experiments.

Article Snippet: Apo2L / TRAIL was obtained from PeproTech (Rocky Hill, NJ), rabbit polyclonal TRAIL neutralizing antibody and control rabbit immunoglobulin from ProSci (Poway, CA).

Techniques: Construct, Transduction, Virus, Clone Assay, Selection, TUNEL Assay, Expressing, Western Blot, Control

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Proteomic analysis of FF samples revealed extracellular matrix (ECM) protein - vitronectin as the component responsible for FTE adhesion and spreading A. Experimental workflow of proteomics experiment. B. Venn diagram showing 14 common proteins identified between young and aged FF samples from proteomics analysis. Common proteins are listed with vitronectin (highlighted in red). C. Representative immunoblot for vitronectin expression in 3 young (Y1–Y3) and 3 aged (A1-A3) FF samples. FF samples (5 μg) and recombinant vitronectin protein (0.01 μg, 0.1 μg, 1 μg) diluted with lysis buffer were used for immunoblotting. Arrow represents the band analyzed for vitronectin. Ponceau staining was used for loading control.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques: Western Blot, Expressing, Recombinant, Lysis, Staining, Control

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Vitronectin in FF aids in FTE adhesion and spreading A. Representative brightfield images of FT190 cells seeded on ULA plates coated with FF sample (400 μl) and recombinant vitronectin protein (1 μg/well) for 4 h. Wells were washed with 1X PBS and FT190 cells were seeded on the coated plates. Images were acquired after 24 h. Scale bar = 200 μm. B. Cell proliferation was measured using an SRB assay to measure cell viability of FT190 cells on FF and vitronectin coated ULA plates. C. Representative Z stack images acquired by confocal microscopy of FT190 spheroids (labelled with Cell tracker 594) on NOF151 cells (labelled with Cell tracker 488) with FF samples and recombinant vitronectin protein (1 μg) (24 h). Scale bar = 200 μm. D. A maximum intensity projection re-construction of a three-dimensional FTE spheroid (labelled with Cell tracker 594) optical data stack over the surface of NOF151 cells (labelled with Cell tracker 488) treated with FF samples (Y1, A1) and vitronectin (1 μg). Spheroids were imaged with a 10× objective and re-constructed using the Imaris software. E. Representative images acquired by confocal microscopy showing a side and top projection of the FT190 spheroids on NOF151 cells with FF samples and recombinant vitronectin protein (1 μg) (24 h). Scale bar = 100 μm. F. Spheroid area (with and without FF/vitronectin) was determined by quantification of red fluorescent intensity using a polygon area measurement tool of the Imaris software.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques: Recombinant, Sulforhodamine B Assay, Confocal Microscopy, Construct, Software

Journal: Heliyon

Article Title: Follicular fluid aids cell adhesion, spreading in an age independent manner and shows an age-dependent effect on DNA damage in fallopian tube epithelial cells

doi: 10.1016/j.heliyon.2024.e27336

Figure Lengend Snippet: Primary antibodies.

Article Snippet: Anti-rabbit vitronectin , Proteintech 15833-1-AP , 1:1000.

Techniques:

Journal: Cancer Biology & Therapy

Article Title: MOICS, a novel classier deciphering immune heterogeneity and aid precise management of clear cell renal cell carcinoma at multiomics level

doi: 10.1080/15384047.2024.2345977

Figure Lengend Snippet: The hub role of SAA4 in ccRCC.

Article Snippet: Human recombinant protein of SAA4 (HY-P71277) was purchased from MedChemExpress, which was added to cell culture medium of 786–0 and ACHN.

Techniques: