anti ube2e3  (Bioss)

Journal: PeerJ

Article Title: UBE2E3 regulates cellular senescence and osteogenic differentiation of BMSCs during aging

doi: 10.7717/peerj.12253

Figure Lengend Snippet: (A) The expression of UBE2E3 in 7862 normal tissues of GTEx. (B) The correlation analysis of UBE2E3 with osteogenesis related genes in GTEx. (C) The correlation analysis of COL1A1 and UBE2E3 in GTEx. (D, E) Heatmap of single cell sequencing of rat BMSCs. (F) UBE2E3 expression of human BMSCs from GSE35956 and GSE35958 . (G) qRT-PCR analyses of UBE2E3 levels in old mouse BMSCs and young mouse BMSCs. (H) Western blot analyses of UBE2E3 levels in old mouse BMSCs and young mouse BMSCs. (I) qRT-PCR analyses of UBE2E3 levels in increasing numbers of passages of mice BMSCs. (J) Western blot analyses of UBE2E3 levels in increasing numbers of passages of mice BMSCs. Error bars showed standard deviation. * P < 0.05, *** P < 0.001.

Article Snippet: Next, membranes were blocked at room temperature with 5% defatted milk for 1 h. Then the membrane was incubated with anti- UBE2E3 (bs-8352R, 1:500; Bioss), anti- GAPDH (10494-1-AP,1:5000; Proteintech) overnight at 4 °C.

Techniques: Expressing, Sequencing, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: PeerJ

Article Title: UBE2E3 regulates cellular senescence and osteogenic differentiation of BMSCs during aging

doi: 10.7717/peerj.12253

Figure Lengend Snippet: (A) Knockdown efficiency of UBE2E3 was verified by qRT-PCR. (B) Knockdown efficiency of UBE2E3 was verified by western blot. (C) β-Gal staining and quantification of senescent BMSCs in siUBE2E3 group and siNC group. (D) The relative mRNA expression of p16 and p21 between siUBE2E3 group and siNC group. (E) ALP and ARS staining results in siUBE2E3 group and siNC group, with the quantitative analysis. (F) The relative mRNA expression of RUN2, ALP, SP7 and BGLAP between siUBE2E3 group and siNC group. Error bars showed standard deviation. * P < 0.05, ** P < 0.01.

Article Snippet: Next, membranes were blocked at room temperature with 5% defatted milk for 1 h. Then the membrane was incubated with anti- UBE2E3 (bs-8352R, 1:500; Bioss), anti- GAPDH (10494-1-AP,1:5000; Proteintech) overnight at 4 °C.

Techniques: Quantitative RT-PCR, Western Blot, Staining, Expressing, Standard Deviation

Journal: PeerJ

Article Title: UBE2E3 regulates cellular senescence and osteogenic differentiation of BMSCs during aging

doi: 10.7717/peerj.12253

Figure Lengend Snippet: (A) Overexpression efficiency of UBE2E3 was verified by qRT-PCR. (B) Overexpression efficiency of UBE2E3 was verified by western blot. (C) β-Gal staining and quantification of senescent BMSCs in UBE2E3 overexpression group and the control group. (D) The relative mRNA expression of p16 and p21 between UBE2E3 overexpression group and the control group. (E) ALP and ARS staining results in UBE2E3 overexpression group and the control group, with the quantitative analysis. (F) The relative mRNA expression of RUN2, ALP, SP7 and BGLAP between UBE2E3 overexpression group and the control group. Error bars showed standard deviation. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Next, membranes were blocked at room temperature with 5% defatted milk for 1 h. Then the membrane was incubated with anti- UBE2E3 (bs-8352R, 1:500; Bioss), anti- GAPDH (10494-1-AP,1:5000; Proteintech) overnight at 4 °C.

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Staining, Expressing, Standard Deviation

Journal: PeerJ

Article Title: UBE2E3 regulates cellular senescence and osteogenic differentiation of BMSCs during aging

doi: 10.7717/peerj.12253

Figure Lengend Snippet: (A) The correlation of NQO1 and UBE2E3 in GSE35956 . (B) The correlation analysis of GCLC and UBE2E3 in GSE35956 . (C) The correlation analysis of GCLC and UBE2E3 in GSE35958 . (D) The correlation of GCLM and UBE2E3 in GSE35958 . (E) Representative images of BMSCs treated with siNC and siUBE2E3, and stained with DAPI and antibody against Nrf2. Scale bar, 50 µm. (F) NQO1, GCLC and GCLM expression between siUBE2E3 group and siNC group as analyzed by qRT-PCR. Error bars showed standard deviation. * P < 0.05, *** P < 0.001.

Article Snippet: Next, membranes were blocked at room temperature with 5% defatted milk for 1 h. Then the membrane was incubated with anti- UBE2E3 (bs-8352R, 1:500; Bioss), anti- GAPDH (10494-1-AP,1:5000; Proteintech) overnight at 4 °C.

Techniques: Staining, Expressing, Quantitative RT-PCR, Standard Deviation

Journal: PeerJ

Article Title: UBE2E3 regulates cellular senescence and osteogenic differentiation of BMSCs during aging

doi: 10.7717/peerj.12253

Figure Lengend Snippet: High expression of UBE2E3 contributes to nuclear translation of Nrf2 and its high activity. With age, the expression of UBE2E3 decreases and nuclear accumulation of Nrf2 and its activity are restrained, leading to cell senescence and reduced osteogenic differentiation potential of BMSCs.

Article Snippet: Next, membranes were blocked at room temperature with 5% defatted milk for 1 h. Then the membrane was incubated with anti- UBE2E3 (bs-8352R, 1:500; Bioss), anti- GAPDH (10494-1-AP,1:5000; Proteintech) overnight at 4 °C.

Techniques: Expressing, Activity Assay