enzyme jak2  (Carna Inc)

Journal: International Journal of Molecular Sciences

Article Title: Identification of Hit Compounds Using Artificial Intelligence for the Management of Allergic Diseases

doi: 10.3390/ijms25042280

Figure Lengend Snippet: The hit compounds exhibited non-toxicity in both BEAS-2B and Jurkat cells, while restoring the suppressed JAK2 activity induced by SOCS3. ( A ) At concentrations up to 100 µM, compound 5 demonstrated no significant decrease in cell viability in BEAS-2B, but at 800 µM, the viability dropped below 50%. ( B ) At concentrations up to 100 µM, compound 8 demonstrated no significant decrease in cell viability in BEAS-2B, but at 800 µM, the viability dropped below 50%. ( C ) At concentrations up to 100 µM, compound 5 demonstrated no significant decrease in cell viability in Jurkat cell, but at 800 µM, the viability dropped below 10%. ( D ) At concentrations up to 100 µM, compound 8 demonstrated no significant decrease in cell viability in Jurkat cell, but at 800 µM, the viability dropped below 10%. ( E ) When the concentration of SOCS3 reached 5.2 µM, the residual activity of JAK2 was 10%. ( F ) When 100 µM of hit compounds 5 or 8 was added, the activity of JAK2, which was reduced from SOCS3 to approximately 25%, recovered to >50%.

Article Snippet: JAK2 enzyme (2.5 ng/µL) was added to the tube and the mixture was allowed to react for 45 min. A Spectramax ® i3x Multi-Mode Microplate Reader (Molecular Devices, San Jose, CA, USA) was used to measure the luminescence, and Softmax ® Pro software version 7.0.2 was used to analyze the results.

Techniques: Activity Assay, Concentration Assay

Journal: bioRxiv

Article Title: Regulation of beta-amyloid production in neurons by astrocyte-derived cholesterol

doi: 10.1101/2020.06.18.159632

Figure Lengend Snippet: (a) Astrocyte-neuron co-cultures are grown from SREBP2 flox/flox Aldh1L1-Cre ERT2 embryos. Astrocyte SREBP2 knockout was induced by application of 4-hydroxytamoxifen (TMX) to cultures. Following knockout, cells were immunostained for the cholesterol synthesis enzyme FDFT1 and the astrocyte marker GFAP and imaged. (b) Mixed primary cultures of neurons and astrocytes derived from SREBP2 flox/flox x Aldh1L1-Cre and Cre negative littermates were treated with 4-hydroxytomoxifen (TMX) to delete SREBP2. APP lipid raft localization in cortical neurons is lost when SREBP2 is knocked out of astrocytes. TMX does not change APP localization in Cre negative cells.

Article Snippet: SREBP2 knockout was confirmed by measuring the downstream enzyme FDFT1 (Abcam, #ab195046, Alexa Fluor 488 secondary Thermo Fisher Scientific #A32766) pTau was imaged in brain sections using the mouse monoclonal AT180 antibody from Thermo Fisher Scientific (#MN1040).

Techniques: Knock-Out, Marker, Derivative Assay