Journal: Cell Reports Medicine

Article Title: Innate cell markers that predict anti-HIV neutralizing antibody titers in vaccinated macaques

doi: 10.1016/j.xcrm.2022.100751

Figure Lengend Snippet:

Article Snippet: Cell-IDTM Intercalator-Rh—2000 μM , Fluidigm , 201103B.

Techniques: Purification, Affinity Purification, Recombinant, Adjuvant, Emulsion, Software

Journal: Cell cycle (Georgetown, Tex.)

Article Title: The reciprocal interaction between autophagic dysfunction and ER stress in adipose insulin resistance.

doi: 10.4161/cc.27406

Figure Lengend Snippet: Figure 6. Osteocalcin inhibits tunicamycin-induced autophagy and ER stress, and improves insulin signaling in adipocytes via NFκB–p65-dependent mechanisms. Cells were pretreated with 5 μg/ml Tun and simultaneously incubated with or without 5 ng/ml of osteocalcin for 4 h. (A) The effect of osteocalcin on ER stress was determinate by the levels of p-PERK and p-eIF2α by immunoblotting. (B) For insulin signaling, cells were stimulated with 50 nM of insulin for 10 min. Insulin receptor signaling was detected by the phosphorylation of IRS-1 and Akt in adipocytes. (C) Atg7, LC3 were examined by immunoblotting, the right is quantification of Atg7 expression by osteocalcin. (D) The function of ocsteocalcin on mTOR activation was detected. All gels are representative of triplicate experiments. (E) The effect of osteocalcin on autophagy and ER stress is in dependence of NFκB. Protein expression of NFκB-p65 in adipocytes transfected with 100 nM NFκB-p65 siRNA. (F) Adipocytes were cultured in the presence or absence of osteocalcin (Ocn) with or without 1 μM PDTC (an NFκB inhibitor) and 100 nM NFκB-p65 siRNA. Protein expression of Atg7 and p62, and the phosphorylation of PERK and IRS-1 in adipocytes. The results are representative of at least triplicate experiments.

Article Snippet: Cells were transfected with a silencing RNA (siRNA) targeted for human Atg7 (Santa Cruz, sc-41447) and human IRE1α synthesized by IDT: AGACAGAGGC CAAGAGCAAA using Lipofectamine 2000 (Invitrogen).

Techniques: Incubation, Western Blot, Phospho-proteomics, Expressing, Activation Assay, Transfection, Cell Culture