Journal: Scientific Reports

Article Title: Effects of fetuin-A-containing calciprotein particles on posttranslational modifications of fetuin-A in HepG2 cells

doi: 10.1038/s41598-021-86881-0

Figure Lengend Snippet: Effects of secondary CPP on expression of fetuin-A mRNA in HepG2 cells. ( A ) HepG2 cells were treated with different concentrations (0, 25, 50, and 100 μg/mL) of synthetic secondary CPP for 24 h. ( B ) HepG2 cells were treated with 100 μg/mL secondary CPP for 0, 6, 12, 24 h. Total RNA was isolated from cells and fetuin-A/ahsg mRNA expression was determined using quantitative real-time PCR. Levels of fetuin-A mRNA were calculated using the comparative cycle threshold method, with 18S ribosomal RNA as the endogenous reference. Fetuin-A mRNA levels without secondary CPP ( A ) or at 0 h ( B ) were defined as 100% (control). Data are shown as mean ± SD from at least three independent experiments. *P < 0.05 vs. control.

Article Snippet: Primers for human fetuin-A (ahsg) (Hs00155659) and 18S ribosomal RNA (4319413E) were purchased from Applied Biosystems (TaqMan Gene Expression Assays).

Techniques: Expressing, Isolation, Real-time Polymerase Chain Reaction, Control

Journal: International Journal of Molecular Sciences

Article Title: Annexins A2, A6 and Fetuin-A Affect the Process of Mineralization in Vesicles Derived from Human Osteoblastic hFOB 1.19 and Osteosarcoma Saos-2 Cells

doi: 10.3390/ijms22083993

Figure Lengend Snippet: Co-localization of AnxA6, AnxA2, or fetuin-A (FetuA) with TNAP in hFOB 1.19 ( A , C , E ) and Saos-2 ( B , D , F ) cells in resting conditions (R) or after seven-day stimulation with AA and β-GP (S). The cells were incubated with appropriate antibodies: mouse monoclonal anti-AnxA6 ( A , B ), mouse monoclonal anti-AnxA2 ( C , D ), or mouse monoclonal anti-FetuA ( E , F ), all followed by goat anti-mouse IgG-FITC (green); rabbit polyclonal anti-TNAP followed by goat anti-rabbit IgG-TRITC (red) and DAPI for nuclei (blue) and observed under an Axio Observer.Z1 FM (Carl Zeiss, Poznan, Poland) with Phase contrast and appropriate fluorescent filters, magnification 630 x. Arrowheads indicate protein accumulation in vesicular and/or cluster structures. The yellow color and arrows on the merge images indicate AnxA6 ( A , B ), AnxA2 ( C , D ), or FetuA ( E , F ) co-localization with TNAP. Results of a typical experiment are presented.

Article Snippet: Then, cells were incubated with mouse monoclonal anti-annexin A6 (AnxA6; 1:100, v/v ; BD Transduction Laboratories, Warsaw, Poland), mouse monoclonal anti-annexin A2 (AnxA2; 1:100, v/v ; BD Transduction Laboratories, Warsaw, Poland), mouse monoclonal anti-fetuin-A (FetuA; 1:100, v/v ; Abcam, Cambridge, UK), or rabbit polyclonal anti-TNAP (TNAP; 1:100, v/v ; Abcam, Cambridge, UK) primary antibodies diluted in TBS containing 0.5% FBS ( v/v ) and 0.05% Tween-20 ( v/v ).

Techniques: Incubation