Journal: PLOS One

Article Title: FOXO1 transcription factor modulates airway epithelial responses to viral infection

doi: 10.1371/journal.pone.0345169

Figure Lengend Snippet: A) RT-qPCR showed increased TLR3 mRNA expression for BEAS-2B transfected with a CA-FOXO1 plasmid compared to vector control (cells transfected with an empty plasmid); GAPDH was used as a housekeeping gene (n = 6). Representative Western blot (B) and densitometry analysis (C) of TLR3 expression for BEAS-2B transfected with CA-FOXO1 plasmid compared to vector control, β-actin was used as a loading control (n = 6). Statistical Analysis with t-test, **p < 0.01. D + E) Immunofluorescence staining for BEAS-2B transduced with CA-FOXO1 shows increased FOXO1 protein in the nucleus. FOXO1 (red) was detected using an anti-FOXO1 antibody with a red-fluorescent secondary antibody, F-actin (green) with phalloidin, and nuclei (blue) with DAPI. Images were taken with an Olympus IX81 epifluorescence microscope using a 20X objective lens. Volocity Analysis was used to quantify nuclear localization of FOXO1 by measuring the mean fluorescence intensity of FOXO1 staining colocalized with DAPI. For each group 40−60 cells per slide were analyzed. Statistical Analysis was conducted with ANOVA **** p < 0.001. BEAS-2B cells transduced with FOXO1 or scrambled shRNA lentivirus were analyzed by RT-qPCR for DDX58 (RIG-I, F), MAVS (G), and MYD88 (H) mRNA expression at baseline and after Poly(I:C) stimulation (8 h and 24 h). Expression was normalized to GAPDH and expressed relative to unstimulated scrambled controls (n = 3; ANOVA). (I) NHBE cells were infected with SARS-CoV-2 in the presence or absence of a FOXO1 inhibitor. Total RNA was collected 24 h post-infection, and viral RNA levels were quantified by qRT-PCR, normalized to ACTB, and expressed relative to mock-infected cells (n = 3; paired t-test).

Article Snippet: Primary mouse anti-β-actin mAb (Santa Cruz Biotechnology, SC-69679) and IRdye-conjugated donkey anti-mouse IgG (LI-COR, Lincoln, Neb) were used as a loading control.

Techniques: Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Control, Western Blot, Immunofluorescence, Staining, Transduction, Microscopy, Fluorescence, shRNA, Infection

Journal: Bioengineering

Article Title: Photothermolysis with 1550 nm Fractional Laser Promotes Regeneration of Gingival Mucosa

doi: 10.3390/bioengineering12111180

Figure Lengend Snippet: α-smooth muscle actin in intact gingical tissue ( a , b ) areas treated with 70 kJ ( c , d ), 100 kJ ( e , f ) and 130 kJ ( g , h ) lasers six weeks after treatment. Immunohistochemical reaction with antibodies against α-smooth muscle actin with hematoxylin counterstaining, magnification ×100 ( left column ), ×200 ( right column ), arrow—healing laser treatment area, star—neoangiogenesis.

Article Snippet: Four-μm-thick sections of the formalin-fixed-paraffin-embedded tissue samples were deparaffinized, incubated with 3% H 2 O 2 for 10 min, underwent heat induced epitope retrieval (pH 6.0 sodium citrate buffer, 30 min in 80 °C water bath), additionally blocked with Background Block (Cell Marque, Rocklin, CA, USA) and incubated with mouse monoclonal primary antibodies against α-smooth muscle actin (α-SMA) (A2547, Merck, Rahway, NJ, USA, diluted 1:400) and detected by horseradish peroxidase-conjugated secondary goat antibodies (G-21040, Invitrogen, Carlsbad, CA, USA, diluted 1:1000) and diaminobenzidine with hematoxylin counterstaining.

Techniques: Immunohistochemical staining