akt1 inhibitor  (MedChemExpress)

Journal: Biology

Article Title: Revealing the Improving Effect and Molecular Mechanism of L -Clausenamide in Combating the Acute Lung Injury: Insights from Network Pharmacology, Molecular Docking, and In Vitro Validation

doi: 10.3390/biology14070836

Figure Lengend Snippet: The 3D zoom-out and close-up images, along with the 2D molecular interaction images between L -Clausenamide and ( a ) serum albumin (ALB); ( b ) RAC-alpha serine/threonine-protein kinase (AKT1); ( c ) caspase-3 (CASP3); ( d ) heat shock protein 90-alpha (HS90A); ( e ) estrogen receptors (ESR1); ( f ) epidermal growth factor receptor (EGFR); ( g ) matrix metalloproteinase-9 (MMP-9); ( h ) proto-oncogene tyrosine-protein kinase (SRC).

Article Snippet: The AKT1 protein (Cat No. HY- P74421 ) and AKT inhibitor (AKT-IN-1) (Cat No. HY-18296) were purchased from MCE (Monmouth Junction, NJ, USA).

Techniques:

Journal: Biology

Article Title: Revealing the Improving Effect and Molecular Mechanism of L -Clausenamide in Combating the Acute Lung Injury: Insights from Network Pharmacology, Molecular Docking, and In Vitro Validation

doi: 10.3390/biology14070836

Figure Lengend Snippet: The molecular mechanism of L -Clausenamide: ( a , b ) The SPR analysis between AKT1 and L -Clausenamide. ( c ) The Western blot analysis of AKT, p-AKT, Cleaved Caspase-3, and β-actin. ( d ) The relative protein level of p-AKT/AKT. ( e ) The relative protein level of Cleaved Caspase-3/β-actin. All results are presented as mean ± S.D., and the experiments were repeated in triplicate. ns, p > 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: The AKT1 protein (Cat No. HY- P74421 ) and AKT inhibitor (AKT-IN-1) (Cat No. HY-18296) were purchased from MCE (Monmouth Junction, NJ, USA).

Techniques: Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Dentin sialoprotein promotes endothelial differentiation of dental pulp stem cells through DSP aa34–50 –endoglin–AKT1 axis

doi: 10.1016/j.jbc.2025.108380

Figure Lengend Snippet: ANGPTL4 , FLT1 , and THBS4 are upregulated by DSP or DSP aa34–50 in DPSCs through activating AKT1 signaling. A , Gene Ontology (GO) analysis of DSP-treated DPSCs compared with BSA-treated cells. B , heatmap of indicated genes in DSP-treated DPSCs compared with BSA-treated cells. C , the mRNA levels of indicated genes in DPSCs treated by BSA or DSP (300 ng/ml) by RT–qPCR analysis (n = 3). ACTB was used for normalization. The calculation method was delta–delta Ct. D , Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed the upregulated and downregulated signaling pathways in DSP-treated DPSCs compared with control cells. E and F , DPSCs with or without ENG knockdown or pretreated with MK2206 (2 μM, an inhibitor targeting AKT signaling pathway) or not were cultured with indicated proteins (300 ng/ml). The level of the phosphorylated AKT1 (p-AKT1) in DPSCs was shown by WB analysis. MK2206 is a molecular inhibitor of the AKT pathway. ACTIN served as a loading control, and the relative ratios of p-AKT1 versus AKT1, AKT1 versus ACTIN, and ENG versus ACTIN were shown ( E ). The mRNA levels of ANGPTL4 , FLT1 , and THBS4 in DPSCs shown by RT–qPCR analysis (n = 3). ACTB was used for normalization. The calculation method was delta–delta Ct ( F ). The quantification results are represented as mean ± SD ( C and F ). Student's t test for ( C ) and two-way ANOVA with Tukey's post hoc test for ( F ). BSA, bovine serum albumin; DSP, dentin sialoprotein; DPSC, dental pulp stem cell; qPCR, quantitative PCR; WB, Western blot.

Article Snippet: To block AKT1 signaling, cells were pretreated with the specific inhibitor for AKT, MK2206 (2 μM, MedChemExpress; catalog no.: HY-108232).

Techniques: Quantitative RT-PCR, Control, Knockdown, Cell Culture, Real-time Polymerase Chain Reaction, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Dentin sialoprotein promotes endothelial differentiation of dental pulp stem cells through DSP aa34–50 –endoglin–AKT1 axis

doi: 10.1016/j.jbc.2025.108380

Figure Lengend Snippet: AKT1 inhibition suppresses the DSP- and DSP aa34–50 -induced migration and endothelial differentiation of DPSCs in vitro . DPSCs with or without ENG knockdown or pretreated with MK2206 (2 μM) or not were cultured with indicated proteins (300 ng/ml). A , representative images of scratch wound healing assays in DPSCs with indicated treatment. The leading edges of cells at 0 and 24 h were marked by solid lines and dashed lines , respectively. B , quantitative analysis of migration areas in ( A ) (n = 3). C , RT–qPCR analysis showing the mRNA levels of PECAM1 and KDR in DPSCs with indicated protein treatment for 7 days (n = 3). ACTB was used for normalization. The calculation method was delta–delta Ct. D , WB analysis showing the protein levels of CD31 and VEGFR2 in DPSCs with indicated protein treatment for 7 days. The red arrow points to the bands of VEGFR2. ACTIN served as a loading control. Relative ratios of VEGFR2 versus ACTIN and CD31 versus ACTIN were shown. E , representative images of Matrigel angiogenesis assays of DPSCs. Cells with indicated protein treatment for 7 days were collected and seeded onto Matrigel-coated plates. After incubation for 14 h, cells were stained with CFSE (2 μM). F , quantification of the tube lengths, the number of nodes, and the number of junctions in ( E ) (n = 3). The quantification results are represented as mean ± SD ( B , C , and F ). Two-way ANOVA with Tukey's post hoc test ( B , C , and F ). Scale bars represent 200 μm ( A and E ). CFSE, carboxyfluorescein succinimidyl ester; DSP, dentin sialoprotein; DPSC, dental pulp stem cell; ENG, endoglin; qPCR, quantitative PCR; VEGFR2, vascular endothelial growth factor receptor 2.

Article Snippet: To block AKT1 signaling, cells were pretreated with the specific inhibitor for AKT, MK2206 (2 μM, MedChemExpress; catalog no.: HY-108232).

Techniques: Inhibition, Migration, In Vitro, Knockdown, Cell Culture, Quantitative RT-PCR, Control, Incubation, Staining, Real-time Polymerase Chain Reaction

Journal: The Journal of Biological Chemistry

Article Title: Dentin sialoprotein promotes endothelial differentiation of dental pulp stem cells through DSP aa34–50 –endoglin–AKT1 axis

doi: 10.1016/j.jbc.2025.108380

Figure Lengend Snippet: AKT1 mediates the endothelial differentiation of DPSCs initiated by DSP and DSP aa34–50 ex vivo . DPSCs with or without ENG knockdown or pretreated with MK2206 (2 μM) or not followed by treatment with indicated proteins (300 ng/ml). After 7 days, cells were harvested and mixed with Matrigel. The mixtures were injected subcutaneously into the ventral side of mice, and the Matrigel plugs were collected after 7 days. A , representative images of Matrigel plugs containing DPSCs with indicated treatment. Asterisks show blood vessels (n = 3). B , representative immunofluorescent images of CD31 and human mitochondria in Matrigel plugs of ( A ). An antihuman mitochondria antibody was used to mark DPSCs and DPSC-derived cells. C , ratios of CD31+ cells versus total cells and DPSC-derived CD31+ cells versus total CD31+ cells in ( B ) (n = 3). The quantification results are represented as mean ± SD ( C ). Two-way ANOVA with Tukey's post hoc test ( C ). Scale bars represent 2 mm for ( A ) and 50 μm for ( B ). DSP, dentin sialoprotein; DPSC, dental pulp stem cell; ENG, endoglin.

Article Snippet: To block AKT1 signaling, cells were pretreated with the specific inhibitor for AKT, MK2206 (2 μM, MedChemExpress; catalog no.: HY-108232).

Techniques: Ex Vivo, Knockdown, Injection, Derivative Assay

Journal: The Journal of Biological Chemistry

Article Title: Dentin sialoprotein promotes endothelial differentiation of dental pulp stem cells through DSP aa34–50 –endoglin–AKT1 axis

doi: 10.1016/j.jbc.2025.108380

Figure Lengend Snippet: The working model showing a positive role of DSP in the migration and endothelial differentiation of DPSCs through DSP aa34–50 –ENG ZP association and subsequent activation of AKT1 signaling pathway. DSP, dentin sialoprotein; DPSC, dental pulp stem cell; ENG, endoglin.

Article Snippet: To block AKT1 signaling, cells were pretreated with the specific inhibitor for AKT, MK2206 (2 μM, MedChemExpress; catalog no.: HY-108232).

Techniques: Migration, Activation Assay

Journal: The EMBO Journal

Article Title: Immediate early splicing controls translation in activated T-cells and is mediated by hnRNPC2 phosphorylation

doi: 10.1038/s44318-025-00374-8

Figure Lengend Snippet: Reagents and tools table

Article Snippet: MK2206 (AKT1/2/3 inhibitor) , Biomol , SYN-1162-M001.

Techniques: Recombinant, Construct, Sequencing, Reverse Transcription, SYBR Green Assay, Marker, Western Blot, Software, Electroporation, Blocking Assay, Imaging, Mass Spectrometry