Fig. 4

Identification and validation of the target of Tan IIA through ABPP in combination with LC‒MS/MS. A The workflow of the ABPP approach for exploring potential Tan IIA targets. B Competition of protein labelling with IAA-alkynyl after RAW 264.7 cells were treated with Tan IIA. C Identification of potential targets of Tan IIA in RAW 264.7 cells by chemical proteomics. D Tan IIA competes with IAA-alkynyl for binding to the purified recombinant LDHC protein, as determined by an in-gel fluorescence assay. E Molecular docking simulation of Tan IIA binding to the LDHC protein. F Immunofluorescence staining of LDHC (green) and IAA-alkynyl clicked with the red fluorescence dye TAMRA (Bar = 10 μm). G An LDHC scavenger inhibited the expression of osteoclast-specific markers (data are mean ± SD, ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 vs. Control)