Journal: Bioactive materials

Article Title: An Injectable silk-based hydrogel as a novel biomineralization seedbed for critical-sized bone defect regeneration.

doi: 10.1016/j.bioactmat.2024.01.024

Figure Lengend Snippet: Fig. 6. The potential mechanism involved in the osteogenesis induced by the seedbed hydrogel. A) Venn diagram and volcano plot of the upregulated genes influenced by the seedbed hydrogel. B) Relative mRNA expression of SLC13A3 and MKX in BMSCs (n = 3). C) Expression of proteins SLC13A3 and MKX in BMSCs (normalization using GAPDH). D) Schematic diagram of the function of SLC13A3 and its inhibitor. E) Succinate uptake of BMSCs (Scale bar = 100 μm). F) ALP staining of BMSCs with or without inhibition of SLC13A3 (Scale bar = 5 mm). Error bars, mean ± standard deviation, **p < 0.01, ***p < 0.001.

Article Snippet: The PVDF membrane was incubated with primary antibodies against COL1A1 (1:1000 dilution, E8I9Z, Cell Signaling Technology, USA), OPN (1:1000 dilution, AKm2A1, Santa Cruz, USA), SLC13A3 (1:500 dilution, 26184- 1-AP, Proteintech, China), MKX (1:500 dilution, sc-515878, Santa Cruz, USA), Akt and phospho-Akt (1:1000 dilution, 9272S, 9271T, Cell Signaling Technology, USA) and GAPDH (1:1000 dilution, D16H11, Cell Signaling Technology, USA), respectively, and further incubated with appropriate horseradish peroxidase-conjugated secondary antibodies.

Techniques: Expressing, Staining, Inhibition, Standard Deviation

Journal: Bioactive materials

Article Title: An Injectable silk-based hydrogel as a novel biomineralization seedbed for critical-sized bone defect regeneration.

doi: 10.1016/j.bioactmat.2024.01.024

Figure Lengend Snippet: Fig. 7. The intracellular signaling pathway involved in the osteogenesis induced by the seedbed hydrogel. A) KEGG analysis of upregulated genes induced by the biomineralization seedbed. B) GSEA analysis of PI3K-Akt signaling pathway related features in the biomineralization seedbed. C) Western blotting of the activation of Akt and p-Akt in BMSCs (normalization using GAPDH). D) ALP staining of BMSCs with or without inhibition of PI3K-Akt signaling pathway (Scale bar = 5 mm). E) Western blotting of the activation of Akt and p-Akt in BMSCs with a gradient concentration of exogenous succinate or with the biomineralization seedbed. F) Western blotting of the activation of Akt and p-Akt with or without the inhibition of SLC13A3. G) The calcium distribution in mitochondria and cytoplasm in BMSCs (Scale bar = 100 μm). H) Schematic diagram of the regulation of the biomineralization hydrogel seedbed. Error bars, mean ± standard deviation, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The PVDF membrane was incubated with primary antibodies against COL1A1 (1:1000 dilution, E8I9Z, Cell Signaling Technology, USA), OPN (1:1000 dilution, AKm2A1, Santa Cruz, USA), SLC13A3 (1:500 dilution, 26184- 1-AP, Proteintech, China), MKX (1:500 dilution, sc-515878, Santa Cruz, USA), Akt and phospho-Akt (1:1000 dilution, 9272S, 9271T, Cell Signaling Technology, USA) and GAPDH (1:1000 dilution, D16H11, Cell Signaling Technology, USA), respectively, and further incubated with appropriate horseradish peroxidase-conjugated secondary antibodies.

Techniques: Western Blot, Activation Assay, Staining, Inhibition, Concentration Assay, Standard Deviation

Journal: Cell Death & Disease

Article Title: Prognostic impact of the expression/phosphorylation of the BH3-only proteins of the BCL-2 family in glioblastoma multiforme

doi: 10.1038/cddis.2012.150

Figure Lengend Snippet: Characteristics of patients and analyses of overall survival

Article Snippet: The expression of Bad, Bim, pBad and pBim was estimated by ELISAs according to the manufacturer's instructions (PathScan Total Bad Sandwich ELISA Kit (Cell Signaling/Ozyme, St Quentin en Yvelines, France) and Human BIM ELISA Kit (Tebu-Bio, Le Perray en Yvelines, France)) and the pBad S136 and pBim S69 antibodies (Cell Signaling/Ozyme, France).

Techniques:

Journal: Cell Death & Disease

Article Title: Prognostic impact of the expression/phosphorylation of the BH3-only proteins of the BCL-2 family in glioblastoma multiforme

doi: 10.1038/cddis.2012.150

Figure Lengend Snippet: Overall survival versus Bad and/or Bim phosphorylation. ( a ) Estimation of correlation (Pearson's test) between the level of pBad S136 and pBim S69 . ( b ) Kaplan–Meier curves illustrate the absence of significant difference in OS between patients with tumors harboring low level of pBad S136 and patients with high level of pBad S136 . ( c ) Kaplan–Meier curves illustrate the absence of significant difference in OS between patients with tumors harboring low level of pBim S69 and patients with high level of pBim S69 . ( d ) Kaplan–Meier curves illustrate the significant difference in OS between patients with tumors harboring high levels of pBim S69 and pBad S136 and patients devoid of this double hallmark.

Article Snippet: The expression of Bad, Bim, pBad and pBim was estimated by ELISAs according to the manufacturer's instructions (PathScan Total Bad Sandwich ELISA Kit (Cell Signaling/Ozyme, St Quentin en Yvelines, France) and Human BIM ELISA Kit (Tebu-Bio, Le Perray en Yvelines, France)) and the pBad S136 and pBim S69 antibodies (Cell Signaling/Ozyme, France).

Techniques:

Journal: Cell Death & Disease

Article Title: Prognostic impact of the expression/phosphorylation of the BH3-only proteins of the BCL-2 family in glioblastoma multiforme

doi: 10.1038/cddis.2012.150

Figure Lengend Snippet: Presence of pBad S136 and pBim S69 in GBM and kinase activities. ( a ) Graphs and Pearson's test analyze the presence of correlation between the levels of Akt activity and the level of EGFr activity, or the level of pBad S136 or the level of pBim S69 . ( b ) Graphs and Pearson's test analyze the presence of correlation between the levels of Erk activity and the level of EGFr activity, or the level of pBad S136 or the level of pBim S69 .

Article Snippet: The expression of Bad, Bim, pBad and pBim was estimated by ELISAs according to the manufacturer's instructions (PathScan Total Bad Sandwich ELISA Kit (Cell Signaling/Ozyme, St Quentin en Yvelines, France) and Human BIM ELISA Kit (Tebu-Bio, Le Perray en Yvelines, France)) and the pBad S136 and pBim S69 antibodies (Cell Signaling/Ozyme, France).

Techniques: Activity Assay