Journal: Connective tissue research

Article Title: GREM1 inhibits osteogenic differentiation, senescence and BMP transcription of adipose-derived stem cells.

doi: 10.1080/03008207.2020.1736054

Figure Lengend Snippet: Figure 3. GREM1 overexpression inhibited ADSC senescence. (a) SA-β-gal staining showed fewer positive cells in GREM1 over- expressing ADSCs compared to vector control. Scale bar: 100 μm. (b) A quantitative analysis of SA-β-gal positive cells. (c) The relative telomerase activity was measured by the telomerase ELISA kit. (d, e) Real-time RT-PCR results showed that GREM1 overexpression decreased the expressions of P16 (d) and P53 (e). The control gene was GAPDH. The statistical significance was evaluated by Student’s t-test. The error bar was indicated by SD. (n = 3). *p ≤0.05.

Article Snippet: We measured the human telomerase reverse transcriptase levels in the cells by ELISA following the manufacturer’s protocol (Human Telomerase Reverse Transcriptase, TERT ELISA Kit, Cusabio, Wuhan, China).

Techniques: Over Expression, Staining, Expressing, Plasmid Preparation, Control, Activity Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Connective tissue research

Article Title: GREM1 inhibits osteogenic differentiation, senescence and BMP transcription of adipose-derived stem cells.

doi: 10.1080/03008207.2020.1736054

Figure Lengend Snippet: Figure 4. GREM1 knockdown accelerated ADSC senescence. (a) SA-β-gal staining showed an increased number of positive cells in the ADSC-GREM1sh group compared to the control group. Scale bar: 100 μm. (b) The quantitative analysis of SA-β-gal positive cells. (c) Telomerase activity assay results showed that GREM1 knockdown significantly decreased ADSCs telomerase activity. (d, e) Real- time RT-PCR results showed that GREM1 knockdown upregulated the expressions of P16 (d) and P53 (e). The control gene was GAPDH. The statistical significance was evaluated by Student’s t-test. The error bar was indicated by SD. (n = 3). *p ≤0.05.

Article Snippet: We measured the human telomerase reverse transcriptase levels in the cells by ELISA following the manufacturer’s protocol (Human Telomerase Reverse Transcriptase, TERT ELISA Kit, Cusabio, Wuhan, China).

Techniques: Knockdown, Staining, Control, Telomerase Activity Assay, Activity Assay, Quantitative RT-PCR

Journal: bioRxiv

Article Title: The role of Cycloastragenol at the intersection of Nrf-2/ARE, telomerase, and proteasome activity

doi: 10.1101/2022.03.11.483898

Figure Lengend Snippet: (a) HEKn cells were transfected with either oligo duplex specific to Nrf-2, hTERT, or Control siRNA. After 48 h later, cells were harvested, and total protein levels of hTERT, Nrf-2, β1, and β5 were determined by IB. GAPDH was used as a loading control. ( b, c, d ) HEKn cells were transfected with either Nrf-2 or Control siRNA (each of them 2 nM). After 48 h later, cells were treated with indicated concentrations of CA for 24 h. ( b ) The total protein levels of Nrf-2, hTERT, β1, and β5 were determined by IB. ( c ) The telomerase enzyme activity was determined by using ELISA (n:3, *p ≤ 0.05, **p ≤ 0.001, ***p ≤ 0.005). ( d ) The proteasomal β1 and β5 subunit activities were determined by using fluorogenic substrates. The activity data were normalized to cellular total protein level and presented as a fold change compared to control cells treated with DMSO used as a solvent control (n:3, *p ≤ 0.05, **p. ≤ 0.001, ***p ≤ 0.005).

Article Snippet: When HEKn cells reached at confluency of 60-70%, cells were transfected with 2 and 15 nM siRNA against Nrf-2 (Origene, USA, SR321100) and hTERT (Origene, USA, SR322002), respectively.

Techniques: Transfection, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: The role of Cycloastragenol at the intersection of Nrf-2/ARE, telomerase, and proteasome activity

doi: 10.1101/2022.03.11.483898

Figure Lengend Snippet: HEKn cells were transfected with either oligo duplex specific to hTERT or Control siRNA. After 48 h later, cells were treated with CA for 24 h. ( a ) The total protein levels of Nrf-2, hTERT, β1, and β5 were determined by IB. ( b ) Before performing the nuclear Nrf-2 activity, cellular fractionation was performed. The Nrf-2 activity was determined by ELISA. The Nuclear activity data was normalized to nuclear protein level and presented as a fold change compared to control cells treated with DMSO used as solvent control. Error bars are presented as standard deviations (n = 3; *p ≤ 0.05, **p ≤ 0.001, ***p ≤ 0.005). ( c ) The proteasomal β1 and β5 subunit activities were evaluated via fluorogenic substrates. The data was normalized to cellular total protein level and presented as a fold change compared to control cells treated with DMSO used as solvent control. Error bars are presented as standard deviations (n = 3; *p ≤ 0.05, **p ≤ 0.001, ***p ≤ 0.005).

Article Snippet: When HEKn cells reached at confluency of 60-70%, cells were transfected with 2 and 15 nM siRNA against Nrf-2 (Origene, USA, SR321100) and hTERT (Origene, USA, SR322002), respectively.

Techniques: Transfection, Activity Assay, Cell Fractionation, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: The role of Cycloastragenol at the intersection of Nrf-2/ARE, telomerase, and proteasome activity

doi: 10.1101/2022.03.11.483898

Figure Lengend Snippet: ( a, b, c ) HEKn cells were transfected with Nrf-2, hTERT, and Control siRNA. After 48 h later, cells were pre-treated with the desired concentration of CA for 8 h, then treated with 6-OHDA for 16 h. The cell viability was assessed by MTT reagent. The absorbance value of cells treated with DMSO as a solvent control for each experiment was considered 100%, and the viabilities of others were calculated. This assay was performed by triplicate samples. Student’s t-test was used to determine the significance of the differences Error bars are presented as standard deviations (n = 3; *p ≤ 0.05, **p ≤ 0.001, ***p ≤ 0.005).

Article Snippet: When HEKn cells reached at confluency of 60-70%, cells were transfected with 2 and 15 nM siRNA against Nrf-2 (Origene, USA, SR321100) and hTERT (Origene, USA, SR322002), respectively.

Techniques: Transfection, Concentration Assay

Journal: Biotechnology Letters

Article Title: Development of HPV 16,18,31,45 E5 and E7 peptides-based vaccines predicted by immunoinformatics tools

doi: 10.1007/s10529-020-02792-6

Figure Lengend Snippet: a 3D view of “VFIYIPLFVIHTHASF” linear B cell epitope on the E5 peptide construct. b 3D view of “FAAYSSAD” linear B cell epitope on the E7 peptide construct. c 3D view of “A:S1, A:A2, A:F3” discontinuous B cell epitope on the E5 peptide construct. d 3D view of “A:F15, A:A16, A:A17, A:Y18, A:S19, A:S20, A:A21, A:D22” discontinuous B cell epitope on the E7 peptide construct. Yellow bubbles show the position of B cell epitopes on the E5 and E7 peptide constructs

Article Snippet: Tsang et al. analyzed HPV16 E6 and E7 proteins and found six immunogenic epitopes, as well (Tsang et al. ).

Techniques: Construct

Journal: Biotechnology Letters

Article Title: Development of HPV 16,18,31,45 E5 and E7 peptides-based vaccines predicted by immunoinformatics tools

doi: 10.1007/s10529-020-02792-6

Figure Lengend Snippet: Linear and discontinuous B cell epitopes of the designed vaccine constructs

Article Snippet: Tsang et al. analyzed HPV16 E6 and E7 proteins and found six immunogenic epitopes, as well (Tsang et al. ).

Techniques:

Journal: Alzheimer's & Dementia : Translational Research & Clinical Interventions

Article Title: Alzheimer's disease drug development pipeline: 2024

doi: 10.1002/trc2.12465

Figure Lengend Snippet: Agents in Phase 2 Alzheimer's disease drug development (clinicaltrials.gov accessed January 1, 2024).

Article Snippet: Tertomotide , Disease‐modifying biologic , Synaptic plasticity/neuroprotection , Human telomerase reverse transcriptase (hTERT) mimic , NCT05189210 , GemVax & Kael , Oct 2022 , Jul 2023.

Techniques: Mutagenesis, Binding Assay, Activity Assay, Clinical Proteomics, Virus, Reverse Transcription, Plasmid Preparation, Expressing