Journal: Frontiers in Nutrition

Article Title: Maternal docosahexaenoic acid supplementation during lactation improves exercise performance, enhances intestinal glucose absorption and modulates gut microbiota in weaning offspring mice

doi: 10.3389/fnut.2024.1423576

Figure Lengend Snippet: Effect of maternal DHA supplementation during lactation on the mTOR signaling pathway in weaning mice. (A) Representative Western blots of p-mTOR (S2448), total mTOR, p-p70S6 kinase (T389), total p70S6 kinase, p-S6 ribosomal protein (S240/244) and total S6 ribosomal protein, and (B, C, D) the relative band intensities of phosphorylation levels normalized to their respective total proteins in jejunal tissue of mice from different groups. Data were expressed as the mean ± SEM; n = 4 per group. Significance was set at p < 0.05 ( * p < 0.05, ** p < 0.01). L-DHA: 150 mg/(kg body weight day) DHA; H-DHA: 450 mg/(kg body weight day) DHA. DHA, docosahexaenoic acid; mTOR, mammalian target of rapamycin.

Article Snippet: After transfer, the PVDF membrane was blocked for 1 h in 5% fat-free milk and then was, respectively, incubated overnight at 4°C with dilutions of primary antibodies: GLUT2 (A12307, 1:2000, Abclonal, China), SGLT1 (bs-1128R, 1:2000, Bioss, China), S6 ribosomal protein (abs131865, 1:2000, Absin, China), p-Ser240/244-S6 ribosomal protein (ab215214, 1:2000, Abcam, UK), p70S6 kinase (14485-1-AP, 1,1,000, Proteintech, China), p-Thr389-p70S6 kinase (9,234, 1,1,000, Cell Signaling Technology, United States), mammalian target of rapamycin (mTOR, T55306, 1:2000, Abmart, China), p-S2448-mTOR (5,536, 1,1,000, Cell Signaling Technology), β-actin (T40104, 1:4000, Abmart).

Techniques: Western Blot, Phospho-proteomics

Journal: Advanced Science

Article Title: CGRP Enhances the Regeneration of Bone Defects by Regulating Bone Marrow Mesenchymal Stem Cells Through Promoting ANGPTL4 Secretion by Bone Blood Vessels

doi: 10.1002/advs.202522295

Figure Lengend Snippet: CGRP promotes endothelial angiogenesis by activating the FAK–AKT–VEGF signaling pathway. (A) The activity score for 14 signaling pathways (androgen, estrogen, EGFR, hypoxia, JAK‐STAT, MAPK, NFkβ, PI3K, p53, TGFβ, TNFα, TRAIL, VEGF, and WNT) in scRNA‐seq data. (B) PI3K pathway activity in high‐ and low‐CGRP groups. (C) VEGF pathway activity in high‐ and low‐CGRP groups. (D) Enrichment analysis of differentially expressed genes between high and low CGRP groups. (E) GSEA results of the PI3K‐AKT‐mTOR and VEGF signaling pathways. (F–I) Correlation analysis between CGRP expression and Akt1/Akt2/Akt3/Vegfa expression. (J) Experimental workflow for TMT labeling‐based phosphoproteomic analysis of HMEC‐1. (K) Quantitative analysis of phosphorylation sites and phosphoproteins identified by liquid chromatography–mass spectrometry. (L) GO enrichment analysis of common up‐regulated and down‐regulated phosphoproteins using the package clusterProfiler. The top 20 GO terms are displayed. (M) Kinase enrichment analysis was performed. The enriched kinases are depicted as a bubble plot. (N) Western blotting of p‐FAK 397, p‐FAK 576, p‐FAK 925, FAK, p‐AKT, AKT, p‐ERK, ERK, VEGFA, and tubulin in HMEC‐1 cells treated with different concentrations of CGRP. (O) Western blotting of p‐FAK 397, p‐FAK 576, p‐FAK 925, FAK, p‐AKT, AKT, p‐ERK, ERK, VEGFA, and tubulin at different time points in HMEC‐1 cells treated with the same concentration of CGRP. (P) Western blotting of p‐FAK 397, FAK, p‐ERK, ERK, p‐AKT, AKT, VEGFA, and tubulin in HMEC‐1 cells treated with Control group, CGRP group, and FAK inhibitors (PF‐562271) group.

Article Snippet: Staining was carried out using primary antibodies against CD31 (Abcam, ab182981, 1:500), EMCN (Abcam, ab106100, 1:200), VEGFA (Boster Bio, BA0407, 1:200), Runx2 (Abcam, ab192256, 1:500), CGRP (Cell Signaling Technology, #14 959, 1:800), PGP9.5 (Proteintech, 14730‐1‐AP, 1:800), LEPR (Proteintech, 20966‐1‐AP, 1:50), and ANGPTL4 (Proteintech, 18374‐1‐AP, 1:500).

Techniques: Activity Assay, Protein-Protein interactions, Expressing, Labeling, Phospho-proteomics, Liquid Chromatography, Mass Spectrometry, Western Blot, Concentration Assay, Control

Journal: Advanced Science

Article Title: CGRP Enhances the Regeneration of Bone Defects by Regulating Bone Marrow Mesenchymal Stem Cells Through Promoting ANGPTL4 Secretion by Bone Blood Vessels

doi: 10.1002/advs.202522295

Figure Lengend Snippet: Cgrp − / − mice exhibited reduced bone angiogenesis and lower bone formation, and bone mass than WT controls. (A) Agarose gel electrophoresis images of samples from wild type (WT) and Cgrp knockout (Cgrp−/−) mice. (B) Western blotting of CGRP, p‐FAK 397, p‐FAK 576, p‐FAK 925, FAK, p‐AKT, AKT, p‐ERK, ERK, VEGFA, and tubulin in HMEC‐1 cells with WT mice and CGRP−/− knockout mice. (C and D) Flow cytometry dot plot and quantification of CD31highEmcnhigh endothelial cells from WT and Cgrp−/− mice. (E and F) Representative images and quantification of CD31 (green)‐ and Emcn (red)‐stained femora from WT and Cgrp−/− mice. Scale bar, 100 µm. (G and H) Representative images and quantitation of Vefga (green) immunostaining in the femora of WT and Cgrp−/− mice. Scale bar, 100 µm. n = 3 mice in each group. (I–M) Representative micro‐CT images and quantitative micro‐CT analysis of the trabecular bone microarchitecture of WT and Cgrp−/− mice. Scale bar, 1 mm. (N and O) Representative images of hematoxylin–eosin staining in distal femora and quantification of the number of adipocytes related to the tissue area. Scale bar, 100 µm. (P and Q) Representative images and quantitation of Runx2 (green) immunostaining in femora of WT and Cgrp−/− mice. Scale bar, 100 µm. (R and S) Representative images of TRAP staining and quantification of TRAP‐positive cells on the trabecular bone surfaces of WT and Cgrp−/− mice. Scale bar, 100 µm. n = 3 mice in each group. The data are shown as the mean ± standard deviation. * p < 0.05; ** p < 0.01; *** , and p < 0.001 by Student's t ‐test and one‐way analysis of variance (J–M). BV/TV, trabecular bone volume per tissue volume; Tb. N, trabecular number; Tb. Sp, trabecular separation; Tb. Th, trabecular thickness.

Article Snippet: Staining was carried out using primary antibodies against CD31 (Abcam, ab182981, 1:500), EMCN (Abcam, ab106100, 1:200), VEGFA (Boster Bio, BA0407, 1:200), Runx2 (Abcam, ab192256, 1:500), CGRP (Cell Signaling Technology, #14 959, 1:800), PGP9.5 (Proteintech, 14730‐1‐AP, 1:800), LEPR (Proteintech, 20966‐1‐AP, 1:50), and ANGPTL4 (Proteintech, 18374‐1‐AP, 1:500).

Techniques: Agarose Gel Electrophoresis, Knock-Out, Western Blot, Flow Cytometry, Staining, Quantitation Assay, Immunostaining, Micro-CT, Standard Deviation

Journal: Scientific Reports

Article Title: Integrative bioinformatics and molecular analysis revealed the roles of mTOR/S6K Axis, CASC15 , and miR-30a-3p in laryngeal squamous cell carcinoma

doi: 10.1038/s41598-026-39618-w

Figure Lengend Snippet: Genes/miRNAs/lncRNAs network. The red arrow indicates the direction of the mTOR/S6K axis. Green nodes denote common miRNAs that target mTOR and S6K . The lncRNAs linked to miR-30a-3p . Nodes represent miRNAs and lncRNAs associated with the mTOR/S6K axis. The edges represent predicted regulatory interactions based on TCGA, GEO, and other databases.

Article Snippet: The primary antibodies included mTOR (Cat No: 2971 S, Cell Signaling, 1:1000), S6K (Cat No: M01475-1, Boster Bio, 1:1000), and GAPDH (Cat No: ab8245, Abcam, 1:2500).

Techniques:

Journal: Scientific Reports

Article Title: Integrative bioinformatics and molecular analysis revealed the roles of mTOR/S6K Axis, CASC15 , and miR-30a-3p in laryngeal squamous cell carcinoma

doi: 10.1038/s41598-026-39618-w

Figure Lengend Snippet: The gene expression levels of mTOR , S6K , lncRNA CASC15 , and miR-30a-3p . mTOR (A), S6K (B), and CASC15 (C) genes are upregulated in LSCC tissues as compared to non-tumor tissues. miR-30a-3p (D) expression decreased in LSCC tissues. All data are expressed as mean ± SEM. (* p < 0.05 and **** p < 0.0001).

Article Snippet: The primary antibodies included mTOR (Cat No: 2971 S, Cell Signaling, 1:1000), S6K (Cat No: M01475-1, Boster Bio, 1:1000), and GAPDH (Cat No: ab8245, Abcam, 1:2500).

Techniques: Gene Expression, Expressing

Journal: Scientific Reports

Article Title: Integrative bioinformatics and molecular analysis revealed the roles of mTOR/S6K Axis, CASC15 , and miR-30a-3p in laryngeal squamous cell carcinoma

doi: 10.1038/s41598-026-39618-w

Figure Lengend Snippet: The protein expression levels of mTOR and S6K in the study population. Western blot image, N: normal tissues; T: Tumor ( A ). The mTOR ( B ) and S6K ( C ) protein levels are upregulated in LSCC tissues compared with non-tumor tissues. All data are expressed as mean ± SEM. (**** P < 0.0001).

Article Snippet: The primary antibodies included mTOR (Cat No: 2971 S, Cell Signaling, 1:1000), S6K (Cat No: M01475-1, Boster Bio, 1:1000), and GAPDH (Cat No: ab8245, Abcam, 1:2500).

Techniques: Expressing, Western Blot

Journal: Frontiers in Nutrition

Article Title: Prevention of high-fat/high-sugar diet-induced type 2 diabetes mellitus-associated non-alcoholic fatty liver disease in rats with fermented and raw Rosa roxburghii Tratt (Cili) juice

doi: 10.3389/fnut.2025.1584551

Figure Lengend Snippet: Expression analyses of transcription or protein levels of selected genes. (A) Expression analyses of selected genes by qRT–PCR. The data represent the means ± SDs from six biological replicates with three technical replicates. (B–D) FCJ/RCJ upregulated ABCC3, IDI1, and APOA2 expression in the liver. FCJ and RCJ significantly upregulated hepatic ABCC3 (B) and IDI1 (C) expression, and RCJ significantly upregulated hepatic APOA2 (D) expression in T2DM rats. The data represent the means ± SDs from three biological replicates with three technical replicates. * p < 0.05, *** p < 0.001, **** p < 0.0001.

Article Snippet: The western blotting antibodies used included APOA2 (BM5624, BOSTER, China), IDI1 (A07892-1, BOSTER, China), ABCC3 (DF3874, Affinity, United States), β-tubulin (Solarbio, China), goat anti-rabbit (BS13278, Bioworld, United States), and goat anti-mouse (BS12478, Bioworld, United States) antibodies.

Techniques: Expressing, Quantitative RT-PCR