Journal: Theranostics
Article Title: JARID1D-dependent androgen receptor and JunD signaling activation of osteoclast differentiation inhibits prostate cancer bone metastasis through demethylating H3K4
doi: 10.7150/thno.104135
Figure Lengend Snippet: JARID1D regulates osteoclast differentiation and bone metastasis in PCa. (A) Cell inflammatory cytokine assay to detect the effects of knocking down JARID1D and its enzymatic activity mutants in 22RV1 and DU145 on the expression of 27 cytokines. (B) RT-PCR detection of RANKL expression in DU145 cells with JARID1D knockdown. (C) ELISA detection of RANKL content in the culture supernatant of DU145 cells after JARID1D knockdown. (D) Schematic diagram of primary osteoclast culture. (E) T tTartrate resistant acid phosphatase (TRAP) staining to assess the differentiation capacity of osteoclasts in different treatment groups and quantification results. (F) RT-PCR analysis of osteoclast differentiation-related gene expression across various treatment groups. (G) Representative TRAP staining images and quantification results of osteoclast differentiation induced by RAW264.7 cell in different treatment groups. (H) RT-PCR analysis of osteoclast differentiation-related gene expression across various treatment groups; (I) Bioluminescence image (left) and radiography image (right) of DU145-NC and DU145 Sh-JARID1D cells intracardially injected into nude mice. (J) Micro-CT image of tibia of hindlimb of representative nude mouse and cross section of tibia of hindlimb of representative nude mouse in DU145-NC and DU145 Sh-JARID1D groups. (K-L) Quantitative map of osteolytic area (K) relative bone volume and trabecular thickness (L) of tibia of hindlimb in (H and I) above (n = 3); (M) Survival analysis comparing the intracardiac injection groups of DU145-NC and DU145 Sh-JARID1D in nude mice. (N) Bone metastasis BLI of two groups of nude mice, DU145-NC and DU145 Sh-JARID1D, through intracardiac injection. (O) H&E and TRAP staining of tibia of hind limbs of nude mice in DU145-NC and DU145 Sh-JARID1D groups. Scale bars, 200, 50 (H&E), and 20 μm (TRAP).
Article Snippet: Human RANKL and mouse IL-2, IL-4, IL-6, TNF, IFN-γ, and IL-1β secretion in cell-culture media were quantified using the species-specific enzyme-linked immunosorbent assay (ELISA) from USCN life science, according to the manufacturer's protocols.
Techniques: Cytokine Assay, Activity Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Knockdown, Enzyme-linked Immunosorbent Assay, Staining, Injection, Micro-CT
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