Journal: bioRxiv

Article Title: Acid stress modulates metabolo-inflammatory pathways in oral epithelial cells

doi: 10.64898/2026.03.16.711383

Figure Lengend Snippet: Schematic representation of the experimental design to investigate the impact of acid stress on oral epithelial cells (OECs) and Toll-like receptor (TLR) agonist stimulation. pH Conditioning : OECs (80% confluence) were cultured in acidified (pH:=:3.0) or complete growth media (pH:=:8.0) for 24h. Morphometric Analysis ( a ): Brightfield micrographs and a machine-learning based image analysis pipeline were used to assess changes in cellular morphology. TLR Agonist Challenge : Cell cultures were subjected to either 100 ng/mL flagellin (TLR5 agonist) or 1 mg/mL Pam3CSK4 (TLR2/1 agonist) for 2-, 6-, or 24h. Molecular profiling ( b ): was conducted on OEC RNA collected after 6h TLR agonist challenge using NanoString® nCounter® technology followed by pathway analysis using the Gene Ontology knowledgebase. A TGF- β ELISA ( c ) was performed on OEC supernatants collected 2-, 6-, and 24h post TLR agonist stimulation. Figure made with Biorender.com

Article Snippet: Low-passage mixed-donor, Primary Human Oral Epithelial Cells (Cat# 36063-01, Celprogen, Inc., Torrance, USA), were thawed from liquid nitrogen, plated on poly-L-lysine-coated T-75 cell culture flasks (Sigma-Aldrich Co., St. Louis, USA; Corning Inc., Durham, USA) and maintained in Human OEC Culture Complete Growth Media (Celprogen, Inc.) containing serum and antibiotics at 37°C and 5% CO 2 for two passages prior to experimentation according to manufacturer recommendations.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Advanced Healthcare Materials

Article Title: Nitric Oxide‐Releasing Catheters with Phenol‐Amine Catalytic Coatings for Improved Anti‐Inflammatory Performance

doi: 10.1002/adhm.202500457

Figure Lengend Snippet: a) HCASMCs viability, b) number of cells, and c) endogenous NO generation after incubation with uncoated and coated catheter segments compared to the blank group, measured using the Live/Dead assay, Hoechst staining, and DAF‐FM diacetate, respectively, at i) 48 h and ii) 72 h. Statistical significance relative to control tests was calculated using one‐way ANOVA, ns = not significant, * p < 0.1, ** p < 0.01, **** p < 0.0001. n = 6; error bars represent standard deviation.

Article Snippet: Human coronary artery smooth muscle cells (HCASMCs) and smooth muscle cell growth medium kit were purchased from Cell Applications.

Techniques: Incubation, Live Dead Assay, Staining, Control, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Expression of Alternative Splice Variants of 6-Phosphofructo-2-kinase/Fructose-2,6-bisphosphatase-4 in Normoxic and Hypoxic Melanoma Cells

doi: 10.3390/ijms22168848

Figure Lengend Snippet: The response of malignant melanoma cells to low oxygen concentration. ( A ) Upper panel: Malignant melanoma lines, WM115 and WM266-4, were cultured with 100 µM pimonidazole (hypoxyprobe) in normoxic (N) and hypoxic conditions (H) for 16 h. The formation of pimonidazole–protein adducts were detected using Western Blot. Lower panel: Ponceau S stained membrane is shown as an internal control for equal protein loading. ( B ) Upper panels: Melanoma cells were cultured for 16 h in normoxia and hypoxia. Then HIF-1 alpha subunit accumulation was verified using the Western Blot. β-actin is shown as an internal control for equal loading. Lower panels: Densitometry analysis of Western Blot bands intensity normalized to β-actin. Relative densitometry value is the average of four independent experiments. The mean ± SEM is shown. Student’s t -test was used to evaluate the influence of hypoxia on HIF-1 alpha subunit stabilization. * p < 0.05 by Student’s t -test, ** p < 0.01 by Student’s t -test, p -value between 0.05 and 0.1 by Student’s t -test was given as an indication of the trend. ( C ) CAIX and PFKFB4 expression was analyzed by RT-qPCR in both melanoma cell lines under hypoxic and normoxic conditions. Expression data for each transcript was normalized to that for the reference gene TBP. Means ± SEM of at least five independent experiments are presented relative to expression in normoxic controls. The Student t -test was used to evaluate the differences between normoxic and hypoxic expression of CAIX and PFKFB4 . ** p < 0.01 by Student’s t -test. ( D ) Upper panels: Melanoma cells were cultured for 16 h in normoxia and hypoxia. Then CAIX and PFKFB4 expression was verified using the Western Blot. β-actin is shown as an internal control for equal loading. Lower panels: Densitometry analysis of Western Blot bands intensity normalized to β-actin. Each relative densitometry value is the average of at least four independent experiments. The mean ± SEM is shown. Studen’st t -test was used to evaluate the influence of hypoxia on CAIX and PFKFB4 expression. * p < 0.05 by Student’s t -test, ** p < 0.01 by Student’s t -test, p -value between 0.05 and 0.1 by Student’s t -test was given as an indication of the trend.

Article Snippet: Two human melanoma cell lines WM115 and WM266-4 were obtained from Rockland Immunochemicals (Limerick, PA, USA).

Techniques: Concentration Assay, Cell Culture, Western Blot, Staining, Membrane, Control, Expressing, Quantitative RT-PCR

Journal: Immunopharmacology and immunotoxicology

Article Title: Circular RNA circCTNNA1 is downregulated in osteoarthritis and sponges miR-29a to suppress LPS-induced apoptosis of synoviocytes.

doi: 10.1080/08923973.2021.1988103

Figure Lengend Snippet: Figure 2. CircCTNNA1 and miR-29a directly interacted with each other in synoviocytes. The interaction between circCTNNA1 and miR-29a was predicted by IntaRNA2.0 (A) and analyzed using RNA pull-down assay (B). The interaction between circCTNNA1 (mut) and miR-29a was also analyzed by RNA pull-down assay (C). p< .05.

Article Snippet: Synoviocytes (type B, primary cells) derived from an adult OA patient were purchased from Sigma-Aldrich (Cat. 408OA-05A) and cultured in human synoviocyte media (Cell applications) at 37 C in an incubator with 5% CO2 and 95% humidity.

Techniques: Pull Down Assay

Journal: Immunopharmacology and immunotoxicology

Article Title: Circular RNA circCTNNA1 is downregulated in osteoarthritis and sponges miR-29a to suppress LPS-induced apoptosis of synoviocytes.

doi: 10.1080/08923973.2021.1988103

Figure Lengend Snippet: Figure 3. CircCTNNA1 and miR-29a failed to regulate the expression of each other in synoviocytes. To explore the interaction between circCTNNA1 and miR-29a, synoviocytes were transfected with either circCTNNA1 expression vector or miR-29a mimic and the overexpression was confirmed every 24 h until 96 h (B). The effects of circCTNNA1 and miR-29a overexpression on the expression of each other were analyzed by RT-qPCR (B). The correlations between circCTNNA1 and miR- 29a were analyzed by Pearson’s correlation coefficient across both OA (C) and control (D) synovial fluid samples. p < .05.

Article Snippet: Synoviocytes (type B, primary cells) derived from an adult OA patient were purchased from Sigma-Aldrich (Cat. 408OA-05A) and cultured in human synoviocyte media (Cell applications) at 37 C in an incubator with 5% CO2 and 95% humidity.

Techniques: Expressing, Transfection, Plasmid Preparation, Over Expression, Quantitative RT-PCR, Control

Journal: Immunopharmacology and immunotoxicology

Article Title: Circular RNA circCTNNA1 is downregulated in osteoarthritis and sponges miR-29a to suppress LPS-induced apoptosis of synoviocytes.

doi: 10.1080/08923973.2021.1988103

Figure Lengend Snippet: Figure 4. CircCTNNA1 may sponge miR-29a to reduce the apoptosis of synoviocytes induced by LPS. Synoviocytes were treated with LPS at dosages of 0, 2, 4, 6, 8, and 10 lg/ml for 48 h, and the expression of circCTNNA1 (A) and miR-29a (B) was determined by RT-qPCR. The role of circCTNNA1 and miR-29a in regulating the apoptosis of synoviocytes induced by LPS (10 lg/ml for 48 h) was analyzed by cell apoptosis (C). Expression of active (cleaved) caspase-3 in different groups was analyzed by Western blot (D). The role of circCTNNA1 (mut) in cell apoptosis was also analyzed (E). p < .05.

Article Snippet: Synoviocytes (type B, primary cells) derived from an adult OA patient were purchased from Sigma-Aldrich (Cat. 408OA-05A) and cultured in human synoviocyte media (Cell applications) at 37 C in an incubator with 5% CO2 and 95% humidity.

Techniques: Expressing, Quantitative RT-PCR, Western Blot