Journal: Journal of neurochemistry

Article Title: TrkB activation by 7, 8-dihydroxyflavone increases synapse AMPA subunits and ameliorates spatial memory deficits in a mouse model of Alzheimer's disease.

doi: 10.1111/jnc.13432

Figure Lengend Snippet: Fig. 4 7, 8-Dihydroxyflavone (7, 8-DHF) increased the accumulation of both GluA1 and GluA2 subunits at the synapses in Tg2576 mice. (a) Representative blots of GluA1-4 from synaptoneurosomal lysates of hippocampus (n = 8 for each group). (b) 7, 8-DHF increased the levels of GluA1and GluA2, and remained the levels of GluA3 and GluA4 unchanged. (c) Representative blots of GluA1-4 from whole lysates of hippocampus. (d) 7, 8-DHF did not affect the levels of GluA1-4 in whole lysates of hippocampus. (e) Representative blots of Ser818 and Ser831of GluA1, Ser880 of GluA2, and Ser845 of GluA1 from synaptoneurosomal extracts of hippocampus. (f) 7, 8-DHF increased the levels of Ser831, and Ser818, and remained the level of Ser880 and Ser845 unchanged. Error bars show SD in all results. *p < 0.05, **p < 0.01 compared to the wild-type (WT)- vehicle mice. #p < 0.05, ##p < 0.05 compared to Tg-vehicle mice.

Article Snippet: Comparable motor and visual function between various groups were observed in the Table 1 The list of primary antibodies used in this study Product name Host Categories No Companies Concentration AMPA Receptor (GluR1) (D4N9V) Rabbit 13185 Cell Signaling Technology, Beverly, MA, USA 1 : 1000 AMPA Receptor (GluR2) (E1L8U) Rabbit 13607 Cell Signaling Technology 1 : 1000 AMPA Receptor (GluR3) Rabbit 3437 Cell Signaling Technology 1 : 1000 AMPA Receptor (GluR4) (Arg860) Rabbit 3825 Cell Signaling Technology 1 : 1000 Phospho-AMPA Receptor (GluR1) (Ser818) Rabbit Cell Signaling Technology 1 : 1000 Phospho-AMPA Receptor (GluR1) (Ser831) Rabbit A4352 Sigma-Aldrich 1 : 1000 Phospho-AMPA Receptor (GluR1) (Ser845) (D10G5) Rabbit 8084 Cell Signaling Technology 1 : 500 Phospho-AMPA Receptor (GluR2) pS880 Rabbit MABN103 Merck Millipore, Temecula, CA, USA 1 : 200 BDNF Rabbit ab46176 Abcam Cambridage, UK 1 : 100 TrkB Rabbit ab33655 Abcam 1 : 200 Phospho-TrkB (Tyr816) Rabbit Merck Millipore 1 : 100 CaMKII Mouse ab3908 Abcam 1 : 1000 Phospho-Erk1/2 Rabbit 9911 Cell Signaling Technology 1 : 500 Akt Rabbit 9272 Cell Signaling Technology 1 : 500 CREB Rabbit 3360R-100 Biovision, Mountain view, CA, USA 1 : 500 Phospho-CaMKII (Thr286) Mouse 3742 Cell Signaling Technology 1 : 1000 Phospho-Erk1/2 Rabbit 3441-100 BioVision 1 : 1000 Phospho-Akt (Ser473) Rabbit 9271 Cell Signaling Technology 1 : 800 Phospho-CREB (Ser133) Rabbit 9191 Cell Signaling Technology 1 : 500 Amyloid precursor protein Rabbit PA5-16730 Thermo Scientific Pierce, Waltham, MA, USA 1 : 200 Amyloid beta 40 Rabbit PA3-16760 Thermo Scientific Pierce 1 : 500 Amyloid beta 42 Rabbit PA3-16761 Thermo Scientific Pierce 1 : 500 Glyceraldehyde-3-phosphate dehydrogenase Rabbit G2267 Sigma-Aldrich 1 : 10000 © 2015 International Society for Neurochemistry, J. Neurochem. (2016) 136, 620--636 (b)(a) (d)(c) (f)(e) (h)(g) (j)(i) © 2015 International Society for Neurochemistry, J. Neurochem. (2016) 136, 620--636 visible-platform test (data not shown).

Techniques:

Journal: Iranian Journal of Public Health

Article Title: Exosomal ITGB2 Mediates Immune Evasion in Triple-Negative Breast Cancer by Suppressing Dendritic Cell Activation via TLR4

doi: 10.18502/ijph.v54i6.18903

Figure Lengend Snippet: Modulation of TLR4 Expression in DCs by Exosomal ITGB2 from TNBC Cells. (A–B) Flow cytometry analysis of dendritic cells (DCs) induced from TNBC patient PBMCs, showing increased proportions of CD80 and CD83 positive cells post-induction, indicating successful DC culture. (C–D) Effects of exosomes from MD-AMB-231 cells with varied ITGB2 expression levels on TLR4 expression in DCs. Exosomes from MD-AMB-231 cells decreased TLR4 expression, while exosomes from ITGB2-knockdown cells restored it, and exosomes from ITGB2-overexpressing cells further reduced TLR4 expression. (E–F) The effect of exosomal ITGB2 on DC maturation (CD80, CD83) in the presence of LPS was evaluated by FCM

Article Snippet: After SDS-PAGE separation and PVDF membrane transfer, membranes were incubated overnight at 4°C with primary antibodies against ITGB2 (#72607), TLR4 (#14358), and β-actin (#3700), which all were purchased from Cell Signaling Technology.

Techniques: Expressing, Flow Cytometry, Knockdown

Journal: Iranian Journal of Public Health

Article Title: Exosomal ITGB2 Mediates Immune Evasion in Triple-Negative Breast Cancer by Suppressing Dendritic Cell Activation via TLR4

doi: 10.18502/ijph.v54i6.18903

Figure Lengend Snippet: Modulation of Cytokine Secretion in DCs by Exosomal ITGB2 from TNBC Cells. (A–E) Levels of cytokines IL-12, IL-4, TNF-α, TGF-β, and IFN-β in DCs treated with exosomes from MD-AMB-231 cells, showing downregulation in MD-AMB-231 and OE- ITGB2 exosome-treated groups and restoration in sh- ITGB2 exosome-treated group. (F) Schematic representation of the mechanism by which TNBC-derived exosomal ITGB2 regulates TLR4 and inhibits dendritic cell maturation and immune response

Article Snippet: After SDS-PAGE separation and PVDF membrane transfer, membranes were incubated overnight at 4°C with primary antibodies against ITGB2 (#72607), TLR4 (#14358), and β-actin (#3700), which all were purchased from Cell Signaling Technology.

Techniques: Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Evaluating the Effect of Lenvatinib on Sorafenib-Resistant Hepatocellular Carcinoma Cells

doi: 10.3390/ijms222313071

Figure Lengend Snippet: The underlying mechanisms of lenvatinib in sorafenib-resistant HCC cells may be through the FGFR4-ERK signaling pathway. ( A ) Proteins expressed in wild-type and sorafenib-resistant HCC cells. ( B ) Huh-7SR and Hep-3BSR cells were treated with 10 μM sorafenib or lenvatinib for 6 h. Protein expression was analyzed using Western blot. ( C ) Huh-7SR and Hep-3BSR cells were treated with 1 and 5 μM sorafenib or lenvatinib for 6 h, and the levels of FGFR4, p-ERK, and ERK were analyzed using Western blot. C.: control; Sor: sorafenib; Len: lenvatinib. ( D ) A lower FGFR4 level is associated with longer overall survival of HCC patients based on TCGA database analysis ( http://www.oncolnc.org , accessed on 5 October 2021).

Article Snippet: The membranes were blocked with 2% skimmed milk (GE Healthcare) in TBST with 0.1% Tween 20 (cat. No. T9142, Takara Bio Inc., Kusatsu, Shiga, Japan) for 30 min and were then incubated overnight at 4 °C with the following primary antibodies: anti-ß-Actin (#66009-1-lg, Proteintech, dilution 1:5000); anti-LC3A/B (#12741, CST, dilution 1:1000); SQSTM1/p62 (#5114, CST, dilution 1:1000); Caspase-3 (#14220, CST, dilution 1:1000); PARP (#9542, CST, dilution 1:1000); Caspase-7 (#12827, CST, dilution 1:1000); EGFR (#PAI-1110, Thermo Fisher Scientific, dilution 1:1000); p-Akt (ser473) (#4060, CST, dilution 1:2000); Akt (#4685, CST, dilution 1:1000); p-mTOR (SER2448) (#5536, CST, dilution 1:1000); mTOR (#2983, CST, dilution 1:1000); cyclinD1 (#2978, CST, dilution 1:1000); p-ERK1/2 (#4370, CST, dilution 1:1000); ERK1/2 (#4695, CST, dilution 1:1000); p-MEK1/2 (#9154, CST, dilution 1:1000); MEK1/2 (#8727, CST, dilution 1:1000); anti-ß-Actin (#4967, CST, dilution 1:1000); FGFR4 (#8562, CST, dilution 1:1000) in 5% serum (cat. No. 9048-46-8, FUJIFILM Wako, Osaka, Japan).

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Molecular Sciences

Article Title: Evaluating the Effect of Lenvatinib on Sorafenib-Resistant Hepatocellular Carcinoma Cells

doi: 10.3390/ijms222313071

Figure Lengend Snippet: Proposed model for the underlying mechanism of lenvatinib in overcoming sorafenib resistance. Overexpression of EGFR leads to the activation of ERK and Akt signaling to induce sorafenib resistance and promote sorafenib-resistant cell proliferation. The underlying advantage of lenvatinib is the inhibition of FGFR4 compared with sorafenib. Huh-7SR and Hep-3BSR cells have high FGFR4 expression and the FGFR4-ERK signaling pathway is the major pathway to overcome increased EGFR-induced ERK activation. Compared with Huh-7SR cells, Hep-3BSR cells showed poor autophagic responsiveness to lenvatinib, which may contribute to their partial cross-resistance to lenvatinib.

Article Snippet: The membranes were blocked with 2% skimmed milk (GE Healthcare) in TBST with 0.1% Tween 20 (cat. No. T9142, Takara Bio Inc., Kusatsu, Shiga, Japan) for 30 min and were then incubated overnight at 4 °C with the following primary antibodies: anti-ß-Actin (#66009-1-lg, Proteintech, dilution 1:5000); anti-LC3A/B (#12741, CST, dilution 1:1000); SQSTM1/p62 (#5114, CST, dilution 1:1000); Caspase-3 (#14220, CST, dilution 1:1000); PARP (#9542, CST, dilution 1:1000); Caspase-7 (#12827, CST, dilution 1:1000); EGFR (#PAI-1110, Thermo Fisher Scientific, dilution 1:1000); p-Akt (ser473) (#4060, CST, dilution 1:2000); Akt (#4685, CST, dilution 1:1000); p-mTOR (SER2448) (#5536, CST, dilution 1:1000); mTOR (#2983, CST, dilution 1:1000); cyclinD1 (#2978, CST, dilution 1:1000); p-ERK1/2 (#4370, CST, dilution 1:1000); ERK1/2 (#4695, CST, dilution 1:1000); p-MEK1/2 (#9154, CST, dilution 1:1000); MEK1/2 (#8727, CST, dilution 1:1000); anti-ß-Actin (#4967, CST, dilution 1:1000); FGFR4 (#8562, CST, dilution 1:1000) in 5% serum (cat. No. 9048-46-8, FUJIFILM Wako, Osaka, Japan).

Techniques: Over Expression, Activation Assay, Inhibition, Expressing