enzyme linked immunosorbent assay elisa total tgfβ1 levels  (Boster Bio)

Journal: BMC veterinary research

Article Title: Knock down of transforming growth factor beta improves expressions of co-stimulatory molecules, type I interferon-regulated genes, and pro-inflammatory cytokine in PRRSV-inoculated monocyte-derived macrophages.

doi: 10.1186/s12917-023-03760-8

Figure Lengend Snippet: Fig. 1 Optimization and validation of knockdown of porcine TGFβ1 mRNA expression and protein translation by AS ODNs. A) MDMs under bright-field microscopy. B) MDMs uptake of fluorescent-labeled siRNA under immunofluorescent microscopy. C) MDMs uptake of fluorescent-labeled siRNA com plexed with different concentrations of transfection reagent and transfection period. D) Effect of TGFβ1 antisense (AS1-4), sense (S1-4) and scramble (Scr1-4) phosphorothioate-modified ODNs on the expression of TGFβ1 mRNA in MDMs stimulated with a mixture of poly I:C and LPS. Band intensities (Additional file 1) indicate the quality of TGFβ1 knockdown. E) Optimization of TGFβAS1 concentration for TGFβ1 mRNA knockdown on MDMs trans fected with TGFβAS1 (0.5, 1, or 2 µM) and stimulated with a mixture of poly I:C and LPS. Band intensities (Additional file 2) indicate the quality of TGFβ1 knockdown. F) TGFβ1 protein levels in MDMs transfected with TGFβAS1 (2 µM) and stimulated with a mixture of poly I:C and LPS. In all figures, error bars indicate the standard deviation (SD). Mean differences of TGFβ1 gene expression or protein translation among groups were tested by one-way ANOVA, followed by Tukey HSD test. Mean differences of percentages of fluoresced cells among groups at time points were tested by one-way repeated measures ANOVA, followed by Tukey HSD test. Different letters indicate significant differences. P < 0.05 was set as a statistically significant level

Article Snippet: Cell culture supernatants were collected for subsequent determination of TGFβ1 protein levels by enzyme-linked immunosorbent assay (ELISA) (Porcine TGF Beta 1 PicoKineTM ELISA kit, Boster Biological Technology, Pleasanton, CA).

Techniques: Biomarker Discovery, Knockdown, Expressing, Microscopy, Labeling, Transfection, Modification, Concentration Assay, Standard Deviation, Gene Expression

Journal: BMC veterinary research

Article Title: Knock down of transforming growth factor beta improves expressions of co-stimulatory molecules, type I interferon-regulated genes, and pro-inflammatory cytokine in PRRSV-inoculated monocyte-derived macrophages.

doi: 10.1186/s12917-023-03760-8

Figure Lengend Snippet: Fig. 4 Effect of TGFβAS1 on TGFβ1 protein translation in PRRSV-2-inoculated MDMs. MDMs were transfected with TGFβAS1, then inoculated with either cPRRSV-2 or HP-PRRSV-2, and stimulated with a mixture of poly I:C and LPS. MDMs inoculated with cPRRSV-2 or HP-PRRSV-2 and stimulated with a mixture of poly I:C and LPS served as PRRSV-2-inoculated control. MDMs treated with transfection media (Tr. media) and inoculated with cPRRSV-2 or HP-PRRSV-2, then stimulated with a mixture of poly I:C and LPS served as PRRSV-2-inoculated/Tr. media control. MDMs inoculated with mock Ag and stimulated with a mixture of poly I:C and LPS served as mock control. Untreated MDMs receiving culture media in the presence or absence of a mixture of poly I:C and LPS served as positive and negative controls, respectively. Cell culture supernatants were collected for ELISA. Error bars indicate the SD. Mean differences of TGFβ1 protein translation among groups were tested by one-way ANOVA, followed by Tukey HSD test. Different letters indicate significant differences. P < 0.05 was set as a statistically significant level

Article Snippet: Cell culture supernatants were collected for subsequent determination of TGFβ1 protein levels by enzyme-linked immunosorbent assay (ELISA) (Porcine TGF Beta 1 PicoKineTM ELISA kit, Boster Biological Technology, Pleasanton, CA).

Techniques: Transfection, Control, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: BMC veterinary research

Article Title: Knock down of transforming growth factor beta improves expressions of co-stimulatory molecules, type I interferon-regulated genes, and pro-inflammatory cytokine in PRRSV-inoculated monocyte-derived macrophages.

doi: 10.1186/s12917-023-03760-8

Figure Lengend Snippet: Fig. 5 Effect of TGFβ knockdown on PRRSV copy numbers in PRRSV-2-inoculated MDMs. MDMs were transfected with TGFβAS1, then inoculated with either cPRRSV-2 or HP-PRRSV-2 (0 h), and stimulated with a mixture of poly I:C and LPS (48 h). MDMs inoculated with cPRRSV-2 or HP-PRRSV-2 and stimulated with a mixture of poly I:C and LPS served as PRRSV-2-inoculated control. MDMs transfected with Scr1, then inoculated with either cPRRSV-2 or HP-PRRSV-2, and stimulated with a mixture of poly I:C and LPS served as PRRSV-2-inoculated/Scr1 control. MDMs treated with transfection media (Tr. media), then inoculated with cPRRSV-2 or HP-PRRSV-2, and stimulated with a mixture of poly I:C and LPS served as PRRSV-2-inoculated/Tr. media control. MDMs receiving mock Ag plus a mixture of poly I:C and LPS served as uninoculated control. Cell culture supernatants were collected for real-time PCR. The CT values were obtained and PRRSV-2 ORF7 RNA copy numbers were calculated based on the CT standard curve generated from 101-108 copies of recombinant PRRSV-2 ORF7 plasmids. Data were presented in log 10 scale of copy number/ml. Error bars indicate the SD. Mean differences of PRRSV-2 ORF7 RNA copy numbers among groups at time points were tested by one-way repeated measures ANOVA, followed by Tukey HSD. Different superscript letters indicate significant difference. P < 0.05 was set as a statistically significant level

Article Snippet: Cell culture supernatants were collected for subsequent determination of TGFβ1 protein levels by enzyme-linked immunosorbent assay (ELISA) (Porcine TGF Beta 1 PicoKineTM ELISA kit, Boster Biological Technology, Pleasanton, CA).

Techniques: Knockdown, Transfection, Control, Cell Culture, Real-time Polymerase Chain Reaction, Generated, Recombinant

Journal: Physiological Reports

Article Title: Dapagliflozin prevents ERK activation and SGLT2‐dependent endoglin upregulation in a mechanically provoked cardiac injury model

doi: 10.14814/phy2.15990

Figure Lengend Snippet: TGF‐β1 and CD105 demonstrated as CAD biomarkers. (a, b) Bar graphs representing serum levels of TGF‐β1 and CD105 with different CAD severity. (c, d) Receiver operating characteristic (ROC) curve and area under the curve (AUC) for TGF‐β1 and CD105 (AUC = 0.701 and 0.731 respectively). Diagonal line indicates zero predictive value for the model. Data were analyzed with the Mann–Whitney U test. Correlations were assessed using Spearman's rank correlation moment correlation coefficient.

Article Snippet: To analyze CD105 and TGF‐β level, the human CD105 ELISA kit (Boster Biological Technology, CatEK0644) and human TGF‐β assay ELISA kit (Boster Biological Technology, CatEK0513) were used to detect CD105 and TGF‐β level, respectively, in the collected serum.

Techniques: MANN-WHITNEY