Journal: The EMBO Journal

Article Title: PIP2 depletion and altered endocytosis caused by expression of Alzheimer's disease‐protective variant PLCγ2 R522

doi: 10.15252/embj.2020105603

Figure Lengend Snippet: A–C A mass ELISA was used to detect specific PIP species after exposure to anti‐FcγRII/III (2.4G2) with or without 3‐a‐aminocholestane (SHIP 1 inhibitor), SF1670 (PTEN inhibitor) and LY294002 (PI‐3K inhibitor). In M‐MOP cells (left panel) and primary microglia (right panel), PI(4,5)P 2 (A), PI(3,4)P 2 (B) and PIP 3 (C) were detected. All data show the mean ± SD of three independent experiments and were analysed by two‐way ANOVA (log‐transformed data were used in C) with Sidak’s multiple comparison tests performed. * P < 0.05, ** P < 0.01, *** P < 0.001 and **** P < 0.0001 (blue: P522 and red: R522).

Article Snippet: Designated wells were inhibited by pre‐exposure for 2 h with 3‐a‐aminocholestane (20 µM, B‐0341), LY294002 (10 µM, B‐0294) or SF1670 (5 µM, B‐0350) (Echelon Biosciences).

Techniques: Enzyme-linked Immunosorbent Assay, Transformation Assay

Journal: Viruses

Article Title: Human Cytomegalovirus Mediates Unique Monocyte-to-Macrophage Differentiation through the PI3K/SHIP1/Akt Signaling Network

doi: 10.3390/v12060652

Figure Lengend Snippet: HCMV engages SHIP1 to induce monocyte-to-macrophage differentiation. ( A ) Human peripheral blood monocytes were pretreated for 1 h with a SHIP1 inhibitor (3AC) at 10 μM. Cells were mock- or HCMV-infected for 48 h, and the percent of cells positive for M1 (CD80 and CD86) and M2 (CD16 and CD163) macrophage markers was measured by flow cytometry. ( B ) Monocytes were pretreated for 1 h with 3AC at 10 μM, then mock- or HCMV-infected for 48 h. Viability was measured through PI and annexin V staining by flow cytometry. ( A , B ) Results are representative of 3–6 independent experiments using monocytes from different donors.

Article Snippet: For the inhibitor studies, the following reagents were used: MK-2206 2HCL (MK; an Akt inhibitor), BYL-719 (BYL; a p110α inhibitor), TGX-221 (TGX; a p110β inhibitor), and CAL-101 (CAL; a p110δ inhibitor) from Selleckchem (Houston, TX, USA), LY-294002 (LY; a pan-PI3K inhibitor) from Calbiochem (Billerica, MA, USA), 3-α-aminocholestane (3AC; a SHIP1 inhibitor) from Echelon Biosciences (Salt Lake City, UT, USA), z-DEVD-fmk (a caspase 3 inhibitor) from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and z-VAD-fmk (a pan-caspase inhibitor) from Selleckchem (Houston, TX, USA).

Techniques: Infection, Flow Cytometry, Staining