Journal: Molecular medicine (Cambridge, Mass.)

Article Title: Microbially produced imidazole propionate impairs prostate cancer progression through PDZK1.

doi: 10.1186/s10020-025-01073-0

Figure Lengend Snippet: Fig. 7 Schematic representation of the mechanism by which IMP inhibits activation of the PI3K-AKT pathway via PDZK1

Article Snippet: Phospho-PI3K p85 alpha (Tyr607) Antibodies was was purchased from Affinity Biosciences (AF3241).

Techniques: Activation Assay

Journal: Scientific reports

Article Title: Characteristics of different lipid droplet-mitochondrial contacts patterns during lipid droplet metabolism in T2DM-induced MASLD.

doi: 10.1038/s41598-025-87871-2

Figure Lengend Snippet: Fig. 1. Insulin resistance and lipid deposition were observed in the liver of db/db mice. (a) Representative western blot maps of IR protein and original blots are presented in Supplementary File 2 Fig. 1. (b) Protein level of IRβ. (c) Protein level of PI3K. (d) Protein level of P-PI3K. (e) Expression level of p-PI3K/PI3K. (f) Protein level of AKT. (g) Protein level of p-AKT. (h) Expression level of p-AKT/AKT. (i) Representative images of liver tissues stained with hematoxylin and eosin (H&E) and Oil Red O (scale bars = 50 μm) are shown. (j) Fat drop area (%). (k) FFA content in the liver. (l) Hepatic levels of TGs. (m) Liver index. Data are mean ± SEM. n = 3 (2 images analyzed per sample for Oil Red O-stained images). n = 8 per group (western blotting, liver FFA, TG, and liver index analyses). Student’s t-test was performed. *P < 0.05, **P < 0.01.

Article Snippet: The membrane was then washed three times with TBST and incubated with HRP-labeled secondary antibodies at room temperature for 1 h. Details of primary and secondary antibodies used in this study are as follows: Insulin Receptor β (4B8) Rabbit mAb(1:1000, 3025 S, Cell Signaling); PI3 Kinase p85 Alpha Monoclonal antibody(1:20000, 60225-1-Ig, Proteintech); Phospho-PI3K p85 alpha (Tyr607) Antibody(1:1000, #AF3241, Affinity); Akt (pan) (11E7) Rabbit mAb (1:1000, 4685 S, Cell Signaling); Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (1:1000, 4060 S, Cell Signaling); GAPDH Polyclonal antibody(1:20000, 10494-1-AP, Proteintech); VDAC1/2 Polyclonal antibody(1:2000, 10866-1-AP, Proteintech); ATGL Antibody(1:2000, 2138 S, Cell Signaling); ADRP/Perilipin-2 Polyclonal antibody(1:10000, 15294-1- AP, Proteintech); Anti-Mitofusin 2 antibody [NIAR164] (1:5000, ab124773, Abcam); DRP1 Antibody (C-5) (1:1000, sc-271583, SANTA); COX4I1 Monoclonal Antibody(1:2000, E-AB-22002, Cell Signaling); CPT1A Polyclonal antibody(1:1000, 15184-1-AP, Proteintech); Acetyl-CoA Carboxylase (C83B10) Rabbit mAb(1:1000, 3676 S, Cell Signaling); Phospho-Acetyl-CoA Carboxylase (Ser79) (D7D11) Rabbit mAb(1:1000, 11818 S, Cell Signaling); Anti-mouse IgG, HRP-linked Antibody(1:2000, 7076P2, Cell Signaling); Anti-rabbit IgG, HRPlinked Antibody(1:2000, 7074P2, Cell Signaling).

Techniques: Western Blot, Expressing, Staining

Journal: Toxicology and applied pharmacology

Article Title: Bioinformatics and network pharmacology identify promotional effects and potential mechanisms of ethanol on esophageal squamous cell carcinoma and experimental validation.

doi: 10.1016/j.taap.2023.116615

Figure Lengend Snippet: Fig. 6. Ethanol activates the PI3K/AKT signaling pathway. (A) Indicates the expression of PI3K and p-PI3K proteins as well as statistical plots. (B) Indicates the expression of AKT and p-AKT proteins as well as statistical plots. *P < 0.05, **P < 0.01.

Article Snippet: The following antibodies were used: PI3 kinase p85 antibody (Cell Signaling #4292), phosphorylated PI3K p85 alpha (Tyr607) antibody (Affinity #AF3241); Akt1 (D9R8K) rabbit mAb (Cell Signaling #75692), phosphorylated AKT1- T308 rabbit pAb (ABclonal AP0304), and IgG secondary antibody (1:50,000, 611–145-002, ROCKLAND, USA).

Techniques: Expressing

Journal: Cell Biology and Toxicology

Article Title: S100A2 activation promotes interstitial fibrosis in kidneys by FoxO1-mediated epithelial-mesenchymal transition

doi: 10.1007/s10565-024-09929-7

Figure Lengend Snippet: FoxO1 is a potential target of S100A2. ( A ) GO and ( B ) KEGG enrichment analyses for differentially expressed genes in TGF-β1-stimulated HK-2 cells with S100A2 knockdown. BP, CC, MF denote biological processes, cellular components, and molecular functions, respectively. S100A2 knockdown blocks ( C , F ) PI3K and activates ( D , G ) FoxO1. ( E , H ) Overexpression of S100A2 inhibits FoxO1 levels. ( I ) Co-IP demonstrates the interaction between S100A2 and FoxO1. ( J ) IF confirms the co-localization of S100A2 and FoxO1. Scale bar, 25 μm. ( K - M ) Immunoblot analysis of nuclear and cytoplasmic levels of p-FoxO1, FoxO1, p-Smad2/3, Snail1, and S100A2 in TGF-β1-stimulated HK-2 cells. ( N ) The effect of TGF-β1 treatment on the fluorescence intensity of p-FoxO1 in HK2 cells. * P < 0.05, ** P < 0.01, *** P < 0.001. ns, no statistical difference

Article Snippet: After blocking with 5% non-fat milk, the membrane was probed with the relevant primary antibodies overnight including anti-Vimentin (Santa Cruz), E-cadherin (Abcam), α-SMA (Santa Cruz), N-cadherin (Abcam), COL1 (Proteintech), S100A2 (Abcam), Smad2/3 (Abcam), p-Smad2/3 (Thr8, Abcam), p-PI3K (Tyr607, Affinity, USA), PI3K (Proteintech), p-AKT (Ser473, CST), AKT (CST), Sirt1 (Affinity), p-FoxO1 (Ser256, Affinity), FoxO1 (Affinity), Snail1 (Abcam), COL3 (Proteintech), GAPDH (Proteintech), H3 (Proteintech), and β-actin (Affinity).

Techniques: Knockdown, Over Expression, Co-Immunoprecipitation Assay, Western Blot, Fluorescence