Journal: RSC Advances

Article Title: Anti-obesity effects of secondary metabolites from Chrysosplenium flagelliferum : in vitro and in silico studies

doi: 10.1039/d6ra00782a

Figure Lengend Snippet: Effects of compounds 1–12 on lipid accumulation in 3T3-L1 adipocytes. (A) Lipid accumulation under treatment with compound 1–12 at 20 µM. (B and C) Representative Oil Red O staining images compounds 2 and 6, respectively. (D and E) Quantification of lipid accumulation of compounds 2 and 6, respectively. Values are represented as mean ± standard deviation of three repeats. Statistical significance is indicated as * p < 0.05 compared to DMI-treated.

Article Snippet: 3T3-L1 murine preadipocytes (American Type Culture Collection, Manassas, VA, USA) were cultured in DMEM/F-12 medium supplemented with 10% bovine calf serum (Gibco Inc. Grand Island), 100 U per mL penicillin (NY), and 100 μg per mL streptomycin (USA) in a CO 2 incubator at 37 °C with 5% CO 2 .

Techniques: Staining, Standard Deviation

Journal: RSC Advances

Article Title: Anti-obesity effects of secondary metabolites from Chrysosplenium flagelliferum : in vitro and in silico studies

doi: 10.1039/d6ra00782a

Figure Lengend Snippet: Effects of compounds 2 and 6 on adipogenic marker protein expression in 3T3-L1 adipocytes. (A and B) Western blot analysis of adipogenic markers (C/EBPα, PPAR-γ, perilipin-1 and FABP4) in 3T3-L1 adipocytes treated with compounds 2 and 6, respectively.

Article Snippet: 3T3-L1 murine preadipocytes (American Type Culture Collection, Manassas, VA, USA) were cultured in DMEM/F-12 medium supplemented with 10% bovine calf serum (Gibco Inc. Grand Island), 100 U per mL penicillin (NY), and 100 μg per mL streptomycin (USA) in a CO 2 incubator at 37 °C with 5% CO 2 .

Techniques: Marker, Expressing, Western Blot

Journal: bioRxiv

Article Title: Reprogramming insulin receptor activation with a de novo agonist to overcome severe insulin resistance

doi: 10.64898/2026.05.04.722722

Figure Lengend Snippet: (A) Amino acid sequence comparison between insulin and RF-409. Site-1–binding residues are shown in yellow and site-2–binding residues in purple. Disulfide bonds are shown in red. (B) Cryo-EM structure of the human IR fully occupied by insulin (PDB: 6PXV). The two protomers are shown in green and blue. Insulin molecules bound at site-1 are shown in yellow, and those bound at site-2 are shown in purple. Asp707 residues on αCT motifs are highlighted in red. (C) Structural view of insulin (yellow) bound at site-1 of the IR, engaging the L1 domain (green) from one protomer and αCT motif together with the loop of FnIII-1 domain (blue) from the other protomer. Val3 of the insulin A chain (VA3, yellow) interacts with Asp707 in the IR (D707, red). (D) Insulin-interacting residues at IR site-1, shown in yellow. (E) Structural view of insulin (purple) bound to site-2 of the IR, engaging the FnIII-1 domain (blue). (F) Insulin-interacting residues at IR site-2, shown in purple. (G) AlphaFold-predicted structure of the IR bound to two RF-409 molecules. The protomers are shown in green and blue. RF-409 is shown in gray, with site-1 and site-2 components highlighted in yellow and purple, respectively. (H) Alphafold2 model of RF-409 binding to the IR, illustrating a non-canonical binding mode distinct from insulin. The L1 domain (green) of one IR protomer and the FnIII-1 domain (blue) of the other protomer are engaged by RF-409 (gray), with site-1 binding residues highlighted in yellow and the site-2 binding residues in purple. (I) Dose–response analysis showing EC 50 values for IR phosphorylation (pY IR) and downstream signaling (pAKT and pERK). Log-transformed EC 50 values (LogEC 50 ) derived from dose-response curves in differentiated 3T3-L1 adipocytes are shown as mean ± SEM, with corresponding EC 50 values indicated for reference. (J) Lipogenesis assay in primary rat hepatocytes comparing metabolic responses elicited by insulin and RF-409. Data are presented as mean ± SEM; one-way ANOVA; n = 5 independent experiments. (K) Inhibition of gluconeogenesis in primary mouse hepatocytes by insulin and RF-409. Data are presented as mean ± SEM; one-way ANOVA; n = 6 independent experiments.

Article Snippet: 3T3-L1 preadipocytes (ATCC CL-173) were cultured in high-glucose DMEM supplemented with 10% calf serum and 1% penicillin-streptomycin.

Techniques: Sequencing, Comparison, Binding Assay, Cryo-EM Sample Prep, Phospho-proteomics, Transformation Assay, Derivative Assay, Inhibition

Journal: bioRxiv

Article Title: Reprogramming insulin receptor activation with a de novo agonist to overcome severe insulin resistance

doi: 10.64898/2026.05.04.722722

Figure Lengend Snippet: (A) Representative immunoblot of differentiated 3T3-L1 adipocytes fasted for 12 h and treated with the indicated concentrations of insulin or RF-409 for 10 min. (B) Quantification of immunoblot data shown in (A), fit by nonlinear regression. Phosphorylation levels were normalized to total protein and expressed relative to the response to 100 nM insulin. Data are presented as mean ± SEM; n = 3 independent experiments. (C) Representative immunoblot of C2C12-IR cells fasted for 4 h and treated with insulin or RF-409 (10 nM) for 5 min, followed by ligand washout and incubation for the indicated time points.

Article Snippet: 3T3-L1 preadipocytes (ATCC CL-173) were cultured in high-glucose DMEM supplemented with 10% calf serum and 1% penicillin-streptomycin.

Techniques: Western Blot, Phospho-proteomics, Incubation