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TRPV4 mediates marrow adipocyte remodeling in AML
could partly prevent this process (Online Supplementary Figure S5E). Additionally, 4aPDD had no significant effect on the proliferation and apoptosis of FBL-3 cells, as deter- mined by a CCK8 assay (Online Supplementary Figure S5F)
and flow cytometry analysis (Online Supplementary Figure S5G). Thus, these results suggest that this dose of 4aPDD affects adipocytes, rather than directly affecting FBL-3 cells in the co-culture system.
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Figure 4. The PI3K/AKT pathway inhibits the TRPV4 promoter FOXC1. (A) Analysis of TRPV4 upstream transcription factor expression of bone marrow (BM) adipocytes treated with or without recombinant human GDF15 (rhGDF15) for 2 days by RNA sequencing). (B) Different expression of TRPV4-related transcription factor genes fol- lowing rhGDF15 treatment for 2 days. (C) Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) was used to analyze TRPV4 mRNA level after treat- ment with FOXC1-targeted shRNA (shFOXC1) lentivirus for 48 h. (D) Western blot was used to analyze the expression of TRPV4 protein after treatment with shFOXC1 lentivirus for 48 h. (E) Chromatin immunoprecipitation-qPCR analysis of TRPV4 gene level in adipocytes with or without FOXC1 knockdown. (F) BM adipocytes were treat- ed with or without rhGDF15 and PI3K inhibitor (PI3K-IN-1, 2 mM) for 4 days. The levels of FOXC1 and TRPV4 proteins were determined using western blot analysis. β- actin protein was used as an internal control for the western blot analysis. Three independent experiments were performed. **P<0.01, *P<0.05.
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