label-free quantitative proteomic analysis  (LC Sciences)

Journal: Molecular Medicine

Article Title: HLA-F regulates the proliferation of trophoblast via PKM2-dependent glycolysis in the pathogenesis of preeclampsia

doi: 10.1186/s10020-025-01201-w

Figure Lengend Snippet: HLA-F is involved in suppressing the lactylation of PKM2 K305. ( A ) PKM enzyme activity and pyruvate content were significantly lower in placental tissues taken from patients with preeclampsia ( n = 12) than in those from normal term controls ( n = 12). ( B ) Results of Western blot illustrating that the relative protein expression of PKM2 was significantly reduced in placenta from patients with preeclampsia ( n = 12) compared to that of normal term controls ( n = 12). ( C ) Western blot results of the lactylation level of proteins extracted from HLA-F–overexpressing (HLA-F-OE) Jar cells or control-overexpression (Ctrl-OE) Jar cells using anti-lactyllysine (pan-kla). The red box contains the PKM2 protein. ( D ) The proteins from HLA-F-OE Jar cells or Ctrl-OE Jar cells were bound by a PKM2 antibody and lactylation was detected using an anti-lactyllysine antibody. The relative lactylation-PKM2 level was calculated based on an OD value of IP-PKM2-PKM2/IP-PKM2-anti-la. The Co-IP experiment was replicated three times. ( E ) Western blot of Jar cells transfected with a plasmid that overexpressed either PKM2-WT or PKM2-K305R. ( F ) PKM enzyme activity assay of Jar cells transfected with control, PKM2-WT–, or PKM2-K305R–overexpressing plasmid. Three replicates were used for each experiment. Data are expressed as mean ± SD. A line connects the pairs of groups that were significantly different from each other, and T-tests were used for comparison between the two groups. Asterisks indicate significant differences (ns: no significance, * p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: The 4-dimensional label-free quantitative proteomics combined with lactylation analysis (4D-LFQP-LA) was supported by Jingjie PTM BioLabs (Hangzhou, China).

Techniques: Activity Assay, Western Blot, Expressing, Control, Over Expression, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation, Enzyme Activity Assay, Comparison

Journal: Advanced Science

Article Title: Isolation and Comprehensive Analysis of Cochlear Tissue‐Derived Small Extracellular Vesicles

doi: 10.1002/advs.202408964

Figure Lengend Snippet: Proteomics analysis of CDsEVs. A,B) Cluster analysis (A) and Venn diagram (B) of the proteomics data for CDsEVs and cochlear tissue. C) Analysis of the proportion of proteins in CDsEVs within the Vesiclepedia and Exocarta databases. D) The volcano map of differentially expressed proteins. E) Verification of CDsEV‐enriched proteins by WB. F–H) GO (F, G) and KEGG (H) analysis of enriched proteins in CDsEVs. The top cellular components (F) and biological processes (G) are shown.

Article Snippet: Label‐free quantitative proteomics analysis was conducted by BGI Tech Solutions Co., Ltd, Guangdong, China, and the main process was as follows.

Techniques: