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: Scalable longitudinal imaging and transcriptomics of cells in dynamic enclosures

doi: 10.64898/2026.05.05.723030

Figure Lengend Snippet: A: UMAP based on transcriptomic data from primary human preadipocytes differentiated for seven days on a fibronectin-coated flow cell. The colors correspond to different clusters based on transcriptomic analysis. B: Transcriptomic UMAP colored by the lipid accumulation score, defined as the ratio between the BODIPY stain and the nuclear stain in each CCE. The insets show examples of cells that are very close in gene expression space but differ in their lipid content. C: violin plots depicting the distribution of lipid accumulation scores (y axis) across the transcriptomic clusters (x axis). D: actual (x axis) vs predicted (y axis) lipid accumulation scores from the elastic net model. The plot is for the held-out test set (20% of the total data). E: Euler diagram showing the overlap between top-20 differentially expressed genes between transcriptomic clusters (blue) and model-selected predictors of lipid accumulation (pink). F: average Log2 fold-change between clusters (x axis) vs absolute model coefficient (y axis) for the genes selected by the model. The red color indicates genes that are among the top-20 differentially expressed genes between transcriptomic clusters. G: Gene expression UMAP colored by the top-3 positive predictors identified by the model, showing that the expression values of these genes are uniformly distributed across the UMAP based on global transcriptomic differences. H: UMAP based on transcriptomic data for BV2 mouse microglial cells. The colors correspond to different clusters based on transcriptomic analysis. I: transcriptomic UMAP colored by phagocytic activity as measured by pHrodo™ intensity after four hours. J: UMAP based on DINOv2 features, colored by phagocytic activity showing a greater degree of separation between high vs low phagocytic scores, compared to the transcriptomic UMAP in panel H. K: violin plots depicting the distribution of phagocytic scores (y axis) across the transcriptomic clusters (x axis). L: R 2 performance of elastic net models trained on expression-only features, DINOv2-only features or a combination of the two (x axis). The data refers to the held-out test set (20% of the total data). M: actual (x axis) vs predicted (y axis) phagocytic scores from the elastic net model using the combined expression and DINOv2 features. The plot is for the held-out test set (20% of the total data). N: Euler plot showing the overlap between top-20 differentially expressed genes between transcriptomic clusters (blue) and model-selected predictors of phagocytic activity (pink). O: average Log2 fold-change between clusters (x axis) vs absolute models coefficient (y axis) for the genes selected by the expression-only model. The red color indicates genes that are among the top-20 differentially expressed genes between transcriptomic clusters. P: ridge plots displaying the expression level (x axis) of Gpnmb and Clec4e across transcriptomic clusters (x axis). These two genes are among the top positive predictors for the gene expression-based model and have clear mechanistic evidence linking them to the phagocytosis process. However, their expression is very similar across all the transcriptomic clusters.

Article Snippet: Adipogenesis was induced using Adipocytes Differentiation Toolkit for Adipose Derived MSCs and Preadipocytes (ATCC, # PCS-500-050).

Techniques: Staining, Gene Expression, Expressing, Activity Assay

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

Journal: Physiological Reports

Article Title: Adipocyte‐specific FFA2 deletion leads to increased adipose inflammation and is associated with altered intestinal lipid handling in mice

doi: 10.14814/phy2.70875

Figure Lengend Snippet: Loss of adipose FFA2 alters adipose‐intestinal crosstalk, leading to downregulated intestinal fat absorption pathways. (a) Pathway enrichment analysis based on differentially expressed genes from RNA‐Seq of jejunal mucosa collected after 2 weeks of WD + FOS diet treatment at thermoneutrality, n = 4 per group, identifies mild upregulation of immune pathways and cell stress. (b) Pathways involved in fat digestion, absorption, and metabolism are downregulated in Adipoq‐F2‐KO jejunum compared to floxed controls, n = 4 per group. (c) Transwell co‐culture setup: IEC18 intestinal epithelial cells in the top chamber were exposed to conditioned media from differentiated 3 T3‐L1 adipocytes (either empty vector [EV] or FFA2 knockdown [F2KD]). A “rescue” condition (F2KD‐conditioned medium replaced by EV medium halfway through the treatment period) was also tested. (d–f) qPCR of IEC18 cells after conditioned media treatment revealed altered expression of Nfkbiz, Phospho1, and Tns4 in cells treated with F2KD‐conditioned media compared to EV, partly rescued by switching to EV media. Data are mean ± SEM.

Article Snippet: 3 T3‐L1 preadipocytes were obtained from ATCC (CL‐173, ATCC, Manassas, VA, RRID: CVCL_0123) and expanded in culture following the ATCC‐recommended handling instructions (American Type Culture Collection, ).

Techniques: RNA Sequencing, Co-Culture Assay, Plasmid Preparation, Knockdown, Expressing

Journal: Physiological Reports

Article Title: Adipocyte‐specific FFA2 deletion leads to increased adipose inflammation and is associated with altered intestinal lipid handling in mice

doi: 10.14814/phy2.70875

Figure Lengend Snippet: FFA2 knockdown in 3 T3‐L1 cells impairs long‐term lipid storage and maintenance of adipocyte maturity, increases ERK signaling, and sustains pro‐inflammatory gene expression. (a) Stable knockdown validation showing reduced FFA2 mRNA in F2KD vs. empty vector (EV) 3 T3‐L1 cells across multiple passages. (b) Oil Red O staining quantification after standard 10‐day differentiation showing that EV and F2KD cells initially differentiate to store the same amount of lipid. (c) Quantification of Oil Red O staining showing that F2KD cells fail to increase lipid storage in response to acetate treatment. (d) qPCR data showing that FFA2 mRNA expression increases drastically with adipocyte differentiation, likely indicating that this gene is necessary for maintaining adipocyte maturity. (e) Western blot in differentiated adipocytes reveals higher pERK1/2 levels in F2KD cells, indicating elevated MAPK signaling in the absence of FFA2. GAPDH serves as loading control. Complete Western blot with molecular weight markers is shown in Figure . The membrane was sequentially probed with anti‐pERK1/2, stripped and re‐probed with anti‐total ERK1/2, then stripped and re‐probed with anti‐GAPDH. (f–h) qPCR data showing reduced levels of Adipoq , Srebp1 , Plin1 , genes necessary for continued adipocyte maturity, in F2KD cells. (i) qPCR data showing persistently higher CCL2 expression in F2KD adipocytes. (j, k) Oil Red O staining of long‐term differentiation of EV and F2KD cells where EV cells store more lipid over a 30‐day differentiation period and this storage is increased in the presence of acetate. N = 3–4 wells per group for all experiments reported here. Data shown as mean ± SEM; p < 0.05 by t ‐tests or one‐way ANOVA.

Article Snippet: 3 T3‐L1 preadipocytes were obtained from ATCC (CL‐173, ATCC, Manassas, VA, RRID: CVCL_0123) and expanded in culture following the ATCC‐recommended handling instructions (American Type Culture Collection, ).

Techniques: Knockdown, Gene Expression, Biomarker Discovery, Plasmid Preparation, Staining, Expressing, Western Blot, Control, Molecular Weight, Membrane

Journal: Food Science & Nutrition

Article Title: Solvent‐Dependent Biological Activities of Aqueous and Ethanolic Extracts From Edible Insect Larvae

doi: 10.1002/fsn3.71848

Figure Lengend Snippet: Anti‐hypertensive and anti‐adipogenic effects of aqueous and ethanolic larval extracts. (a) Angiotensin‐converting enzyme (ACE) inhibitory activity of larval extracts at the indicated concentrations (20–300 μg/mL), measured using a colorimetric ACE Kit‐WST assay according to the manufacturer's instructions. Results are expressed as percentage inhibition relative to untreated control. (b) Effects of larval extracts on intracellular lipid accumulation in differentiated 3T3‐L1 adipocytes. Cells were induced to differentiate for 8 days in the presence or absence of extracts (100–400 μg/mL), and lipid accumulation was quantified by Oil Red O staining at 520 nm. Untreated differentiated cells (−) served as control group. Data are expressed as mean ± SD from three independent biological experiments ( n = 3), each performed in triplicate. * p < 0.05 versus control.

Article Snippet: Mouse 3T3‐L1 preadipocytes were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) and cultured in DMEM containing 10% bovine calf serum and 1% penicillin–streptomycin under identical incubation conditions.

Techniques: Activity Assay, WST Assay, Inhibition, Control, Staining