Journal: Journal of Dental Sciences

Article Title: Human umbilical cord mesenchymal stem cells secretome and nanoemulsion propolis combination ameliorate osteoclastogenesis in lipopolysaccharide-induced osteolysis in hyperglycemia rats

doi: 10.1016/j.jds.2025.07.003

Figure Lengend Snippet: Effect of NEP and HUCMSCS combination on LPS-induced osteolysis in hyperglycemic conditions in TRAPase and Ctsk serum levels. (A) Concentration of TRAP in serum was measured by ELISA. (B) Concentration of Ctsk in serum was measured by ELISA. A comparison of (A) TRAPase and (B) Ctsk serum levels in all experimental groups. Results are expressed as the mean ± SD. The statistical significance of differences was determined by t-tests and one-way analysis of variance (ANOVA) by Tukey Honest Significant Different (HSD) with significance set at P ≤ 0.05. n = 4 for each group. CTSK, Cathepsin K; HUCMSCS, human umbilical cord mesenchymal stem cells secretome; LPS, lipopolysaccharide; NEP, nanoemulsion propolis extract; PBS, phosphate-buffered saline; SD, standart deviation; TRAP, tartrate-resistant acid phosphatase.

Article Snippet: The steps of the sandwich ELISA method begin with preparation, sample incubation, washing, target antibody incubation, sample washing, analysis, and interpretation according to the instructions of the rat TRAP and Ctsk ELISA kit (Shanghai Korain Biotech Co., Ltd., Shanghai, China).

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison, Saline

Journal: The Febs Journal

Article Title: Biogenesis of hepatitis B virus e antigen is driven by translocon‐associated protein complex and regulated by conserved cysteine residues within its signal peptide sequence

doi: 10.1111/febs.16304

Figure Lengend Snippet: Volcano plots as a result of student's t ‐test comparison of shotgun LC‐MS/MS analysis of proteins co‐immunoprecipitated with HBe‐HA (A) and HBe3CA‐HA (B) expressed in HEPG2‐hNTCP cells. The scatter plot shows differential patterns between the HBe or HBe3CA sample and a negative control sample (CTRL, empty vector) performed in biological triplicates. The blue points in the plot represent Sec61 and TRAP complex subunits, and the red dot represents a bait protein.

Article Snippet: The constructs of individual c‐myc‐tagged human TRAP subunits (α, β, δ) under the CMV promotor were purchased from OriGene Technologies (Rockville, MD, USA; #RC202408, #RC213580, #RC201079, # C201593).

Techniques: Liquid Chromatography with Mass Spectroscopy, Immunoprecipitation, Negative Control, Plasmid Preparation

Journal: The Febs Journal

Article Title: Biogenesis of hepatitis B virus e antigen is driven by translocon‐associated protein complex and regulated by conserved cysteine residues within its signal peptide sequence

doi: 10.1111/febs.16304

Figure Lengend Snippet: Co‐immunoprecipitation of the HBV precore protein with individual TRAP subunits and the TRAP‐depletion effect on precore stability and translocation. (A) HEK 293T cells were co‐transfected with an HA‐tagged HBe‐producing construct (or mock DNA) and plasmids expressing individual c‐myc‐tagged TRAP subunits. The precore protein was immunoprecipitated with anti‐HA magnetic beads and the samples were analysed by western blot using anti c‐myc antibody to detect interacting TRAP proteins. (B) HEK 293T cells were co‐transfected with an HBe‐producing construct and siRNAs targeting either TRAP α or δ genes. The silencing effect was evaluated by western‐blot analysis. The dashed line indicates separated parts of the same membrane. (C) Knockdown of TRAP decreases the efficiency of HBe secretion. Forty‐eight hours after transfection, TRAP‐silenced cells were metabolically labelled for 30 min with 35 S (P) and chased for 4 h (CH) with or without the addition of the proteasome inhibitor MG132. The cells and media were harvested and subjected to immunoprecipitation with anti‐core antibody. The proteins were separated by SDS/PAGE and analysed by autoradiography. (D) The signal intensity of p17 in the medium was quantified and shown relative to the unsilenced sample. The bars represent the averages of three independent experiments, whereas the error bars indicate the standard deviation, * denotes a significance level ≤ 0.05. (E) Representative confocal‐microscopy images of HA‐tagged HBe in HEK 293T cells depleted for the TRAP α subunit visualised by FITC‐conjugated anti‐HA antibody; the ER is stained with anti‐calnexin AlexaFluor647‐conjugated antibody. Calnexin (red), HBe, HBc (green) and DAPI (blue). The scale bar represents 5 μm.

Article Snippet: The constructs of individual c‐myc‐tagged human TRAP subunits (α, β, δ) under the CMV promotor were purchased from OriGene Technologies (Rockville, MD, USA; #RC202408, #RC213580, #RC201079, # C201593).

Techniques: Immunoprecipitation, Translocation Assay, Transfection, Construct, Expressing, Magnetic Beads, Western Blot, Metabolic Labelling, SDS Page, Autoradiography, Standard Deviation, Confocal Microscopy, Staining

Journal: The Febs Journal

Article Title: Biogenesis of hepatitis B virus e antigen is driven by translocon‐associated protein complex and regulated by conserved cysteine residues within its signal peptide sequence

doi: 10.1111/febs.16304

Figure Lengend Snippet: The depletion of TRAP subunits downregulates the extracellular level of HBe but not HBs in HBV‐infected hepatocytes. Biological triplicates of HepG2‐hNTCP cells were first transfected with siRNA oligonucleotides for 24 h, then infected with HBV at a MOI of 1500 VGE/cell for another 96 h before the cells and media were harvested. Secreted HBe (A) and HBs (B) antigens in the media were quantified by ELISA. The error bars represent standard deviations. The asterisks indicate statistically significant differences between the control (siCTRL) and the respective depleted TRAP subunit determined by ANOVA (* P < 0.05; ** P < 0.01). The data are representative of three biological replicates. (C) Knockdown efficiency was verified by RT‐qPCR following total RNA isolation from the cells harvested 3 days after transfection. (D) The cytotoxic effect of the silencing was evaluated by XTT assay 5 days after transfection. TRAP‐subunit knockdown affects HBe secretion for both wt (E) and mutant HBe3CA (F) HBV. HepG2‐hNTCP cells were simultaneously transfected with rcccDNA plasmid coding for either WT or HBe3CA virus and siRNAs targeting individual TRAP subunits. Secreted HBe levels in the media were quantified by ELISA after 5 days of cultivation.

Article Snippet: The constructs of individual c‐myc‐tagged human TRAP subunits (α, β, δ) under the CMV promotor were purchased from OriGene Technologies (Rockville, MD, USA; #RC202408, #RC213580, #RC201079, # C201593).

Techniques: Infection, Transfection, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Isolation, XTT Assay, Mutagenesis, Plasmid Preparation