Journal: Nature

Article Title: Structure of the Insulin Receptor-Insulin Complex by Single Particle CryoEM analysis

doi: 10.1038/nature26153

Figure Lengend Snippet: Human IR ECD binding and kinetic data . Top: Human IR ECD binding data for recombinant human insulin. Data is reported as the geometric mean for 23 replicates; the geometric standard deviation is shown in parentheses. The potency measured with the 125 I-insulin competition assay agrees very well with values from the literature 17 . Bottom: Human IR ECD Binding Kinetic Data for RHI. Data is reported as the geometric mean for three replicates; the geometric standard deviation is shown in parentheses. The SPR data is very consistent with values from the literature 31 .

Article Snippet: Recombinant human Insulin Receptor/CD220 was obtained from R&D systems (catalog # 1544-IR-50/CF, https://www.rndsystems.com/products/recombinant-human-insulin-r-cd220-aa-28-944-protein-cf_1544-ir-cf ), reconstituted in PBS (Phosphate buffered saline, pH=7.4) at 0.3 mg/ml (0.00293 mM), stored in 100 ul aliquots at -80, and used as is (without further purification).

Techniques: Binding Assay, Recombinant, Standard Deviation, Competitive Binding Assay

Journal: Aging (Albany NY)

Article Title: Geniposide-mediated protection against amyloid deposition and behavioral impairment correlates with downregulation of mTOR signaling and enhanced autophagy in a mouse model of Alzheimer's disease

doi: 10.18632/aging.101759

Figure Lengend Snippet: Geniposide treatment decreases mTOR activation markers in brains of APP/PS1 mice. Hippocampal expression of Akt, mTOR, and 4E-BP1, and their respective phosphorylated forms was detected by western blot. The expression of p-Akt ( A ) and p-mTOR ( B ) was enhanced in APP/PS1 mice compared to WT, and geniposide attenuated this increase. The expression of p-4E-BP1 ( C ) in APP/PS1 mice was reduced compared to WT, and geniposide partly restored this decrease. Data are presented as mean ± SEM (n = 6). *** p < 0.001, ** p < 0.01, * p < 0.05 vs. WT; # p < 0.05 vs. APP/PS1 mice (one-way ANOVA, Tukey's Multiple Comparison Test). WT: wild-type mice. GP: geniposide.

Article Snippet: The membranes were blocked in 5% bovine serum albumin in TBST (Tris-buffered saline with 0.05% Tween-20) for 1h, and incubated overnight at 4°C with primary antibodies directed against: Akt (1:1,000), p-Akt (1:2,000), mTOR (1:1,000), p-mTOR (1:1,000), 4E-BP1 (1:1,000), or p-4E-BP1 (1:2,000), followed by incubation at 4°C for 2h with a goat-anti-rabbit IgG-horseradish peroxidase-conjugated secondary antibody (Boster, Wuhan, China).

Techniques: Activation Assay, Expressing, Western Blot, Comparison