Journal: bioRxiv

Article Title: Bio-macromolecular assembly machine for human limb lengthening

doi: 10.1101/2024.08.15.608028

Figure Lengend Snippet: Regulatory mechanism of polarized GP mineralization revealed by proteomics. A) Experimental procedure for LC-MS/MS. The GP samples from 3 biological replicates were divided into five groups: epiphysis (E), GP-epiphysis (GP-E), GP, GP-metaphysis (GP-M) and metaphysis (M) tissue; B-C) Volcano plot showing proteins with differential expression between interfaces and GP tissue/bone tissue (epiphysis and metaphysis) (dotted line showing p-value < 0.05 and two-fold change cut-offs); SPP1, AHSG, ENPP1 and ALPL have been highlighted in the volcano plot; D) Representative images of GP samples immunostained for AHSG (magenta) /calcium (cyan) and SPP1(magenta) /calcium (cyan) at the GP-epiphysis interface; E) Representative images of GP samples immunostained for AHSG/calcium, SPP1/calcium, ENPP1 /calcium and ALPL/calcium at the GP-metaphysis interface, scale bar = 20 μm; F) Summary diagram of the macromolecule-based regulatory mechanism of GP-guided polarized long bone elongation.

Article Snippet: Primary antibodies used were: SPP1 (Santa, sc-21742), AHSG (Proteintech, 66094-1-Ig), ALPL (Proteintech, 11187-1-AP), ENPPI (Abcam, ab223268), PTHrP (Santa, sc-12722), Ki67 (Abcam, ab15580), and COL X (Abcam, ab49945).

Techniques: Liquid Chromatography with Mass Spectroscopy, Quantitative Proteomics

Journal: bioRxiv

Article Title: Bio-macromolecular assembly machine for human limb lengthening

doi: 10.1101/2024.08.15.608028

Figure Lengend Snippet: Schematic summary depicting the concept of “mineralization waves” that govern the growth plate (GP)-guided polarized mineralization process. The inhibitory proteins, SPP1 and AHSG, act as “wave attenuators,” forming a line of defence against mineralization in GP-epiphysis interface. When combining with the mineralization-promoting enzymes, ENPP1 and ALPL at the GP-metaphysis interface, these proteins serve as molecular machinery for “wave amplifiers,” accelerating the formation of ACP and its transformation to HAp and facilitating the progression of the mineralization front.

Article Snippet: Primary antibodies used were: SPP1 (Santa, sc-21742), AHSG (Proteintech, 66094-1-Ig), ALPL (Proteintech, 11187-1-AP), ENPPI (Abcam, ab223268), PTHrP (Santa, sc-12722), Ki67 (Abcam, ab15580), and COL X (Abcam, ab49945).

Techniques: Transformation Assay