torin 2  (MedChemExpress)

Journal: The Journal of Biological Chemistry

Article Title: The E3 ligase β-TRCP1 earmarks OTUD3 for destruction to fine-tune cGAS activation

doi: 10.1016/j.jbc.2026.111461

Figure Lengend Snippet: dsDNA-mediated inactivation of RSK3 stabilizes OTUD3 to promote cGAS activation. AuthorAnonymous, A and C , IB analysis of WCL from BPH1 ( A ) or EA.hy926 cells ( C ) transfected with 5 μg/ml ISD90 or polyethylenimine only for indicated time periods. B and D , qRT-PCR analysis of OTUD3 mRNAs from indicated cells. Error bars represent SD, ∗ p < 0.05 (two-tailed t test), n = 2. E and F , IB analysis of WCL from indicated BPH1 cells transfected with 5 μg/ml ISD90 for indicated time periods. G , IB analysis of WCL from BPH1 cells transfected with 5 μg/ml ISD90 for indicated time periods. Where indicated, cells were pre-treated with 2 μM ERK inhibitor SCH772984 for 1 h. H , IB analysis of WCL from BPH1 cells transfected with 5 μg/ml ISD90 for the indicated time points. Where specified, cells were pre-treated with indicated inhibitors at 2 μM for 1 h prior to transfection. I , IB analysis of WCL from BPH1 cells transfected with 5 μg/ml ISD90 for the indicated time points. Where specified, cells were pretreated with 100 nM MK2206 or 10 μM palbociclib for 1 h prior to transfection. J , IB analysis of WCL from BPH1 cells transfected with 5 μg/ml ISD90 for the indicated time points. Where specified, cells were pre-treated with BKM120 (500 nM), Torin 2 (100 nM), Rapamycin (50 nM), Dasatinib (1 μM), or Sunitinib (10 μM) for 1 h prior to transfection. K and L , IB analysis of WCL from EA.hy926 cells transfected with 5 μg/ml ISD90 for the indicated time points. Where specified, cells were pretreated with 300 nM Torin two for 1 h prior to transfection. M , a cartoon illustration of the proposed model generated by Biorender. ERK, extracellular signal-regulated kinase; IB, immunoblot; qRT-PCR, quantitative real-time PCR; WCL, whole cell lysates.

Article Snippet: BI-D1870 (HY-10510), MLN4924 (HY-70062), AZD6738 (HY-19323), KU-55933 (HY-12016), VE-822 (HY-13902), SCH772984 (HY-50846), MK2206 (HY-108232), Palbociclib (HY-50767), BKM120 (HY-70063), Torin 2 (HY-13002), Rapamycin (HY-10219), Dasatinib (HY-10181), and Sunitinib (HY-10255A) were purchased from MedChemExpress.

Techniques: Activation Assay, Transfection, Quantitative RT-PCR, Two Tailed Test, Generated, Western Blot, Real-time Polymerase Chain Reaction

Journal: Frontiers in Immunology

Article Title: PTEN-mTORC2 signaling module controls antibody isotype selection and antiviral humoral immunity

doi: 10.3389/fimmu.2026.1771230

Figure Lengend Snippet: mTOR signaling negatively regulates IgG1 isotype switching. (A) Schematic of in vitro differentiation of GC-like B cells (iGCBs) and plasmablasts (iPBs). Splenic B cells were co-cultured with feeder cells expressing CD40L and BAFF (CD40LB) in the presence of IL-4 to generate iGCBs, followed by IL-21 to induce iPB differentiation. (B) Time-course immunoblot analysis of mTOR signaling in iGCBs and iPBs. Data shown are representative of two or three independent experiments. (C) Immunoblot analysis of iGCBs treated with rapamycin (Rapa, 5 nM), torin-2 (Torin, 25 nM), or vehicle (Vh). Representative immunoblots from four independent experiments are shown. (D–F) Representative flow cytometry (FACS) plots of iGCBs cultured in the presence of Rapa, Torin, or Vh, gated on viable B cells, with percentages of the indicated populations. (G) Quantification of Ig isotype distribution in iGCBs, presented as pie charts. (H–J) Representative FACS plots of iPBs differentiated from iGCBs treated with Rapa, Torin, or Vh, gated on viable B cells, with percentages of the indicated populations. (K) Quantification of Ig isotype distribution in iPBs. Data in (D–F) and (H–J) are representative of two independent experiments, each performed with three biological replicates. Statistical significance in (G) and (K) was assessed by an unpaired two-tailed Student’s t-test: ***p < 0.001.

Article Snippet: In some experiments, iGCB cultures were treated with 5 nM Rapamycin (Tocris), 25 nM Torin-2 (Tocris), 1 μM MK-2206 (Selleckchem), or 50 μM DAPT (MCE).

Techniques: In Vitro, Cell Culture, Expressing, Western Blot, Flow Cytometry, Two Tailed Test