Journal: EMBO Reports

Article Title: Salt‐inducible kinase induces cytoplasmic histone deacetylase 4 to promote vascular calcification

doi: 10.15252/embr.201643686

Figure Lengend Snippet: Extracellular stimuli such as high extracellular phosphate induce the upregulation of HDAC4 expression. HDAC4 can shuttle between the nucleus and cytoplasm, but phosphorylation by SIK kinase in VSMCs promotes its cytoplasmic accumulation through binding to 14‐3‐3 proteins. SIK was reported to directly bind 14‐3‐3 proteins. In the cytoplasm, HDAC4 binds the actinin‐associated protein ENIGMA. All the components of this cytoskeletal complex are required to promote vascular calcification, as knockdown of either HDAC4 or ENIGMA or the inhibition of SIK results in blunted calcification.

Article Snippet: All primers and probes were ordered from IDT (see Tables and ): table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Species Gene Forward Backward Human HDAC4 GGC CCA CCG GAA TCT GAA C GAA CTC TGG TCA AGG GAA CTG Mouse Actb CTC TGG CTC CTA GCA CCA TGA AGA GTA AAA CGC AGC TCA GTA ACA GTC CG Mouse Runx2 CGG CCC TCC CTG AAC TCT TGC CTG CCT GGG ATC TGT A Mouse Osteocalcin GCA ATA AGG TAG TGA ACA GAC TCC GTT TGT AGG CGG TCT TCA AGC Mouse Sox9 CGA CCC ATG AAC GCC TT GTC TCT TCT CGC TCT CGT TC Mouse Alkp GGC TGG AGA TGG ACA AAT TCC CCG AGT GGT AGT CAC AAT GCC Mouse Spp1 ATC TCA CCA TTC GGA TGA GTC T TGT AGG GAC GAT TGG AGT GAA A Mouse Enigma TGC AAG AAG AAG ATC ACT GGA G CAT TGA AGT CCT TGC CCC CT Mouse Hdac1 AGT CTG TTA CTA CTA CGA CGG G TGA GCA GCA AAT TGT GAG TCA T Mouse Hdac2 GGA GGA GGC TAC ACA ATC CG TCT GGA GTG TTC TGG TTT GTC A Mouse Hdac3 ACC GTG GCG TAT TTC TAC GAC CAG GCG ATG AGG TTT CAT TGG Mouse Hdac4 CAC TGC ATT TCC AGC GAT CC AAG ACG GGG TGG TTG TAG GA Mouse Hdac5 TGC AGC ACG TTT TGC TCC T GAC AGC TCC CCA GTT TTG GT Mouse Hdac6 TCC ACC GGC CAA GAT TCT TC CAG CAC ACT TCT TTC CAC CAC Mouse Hdac7 GAA CTC TTG AGC CCT TGG ACA GGT GTG CTG CTA CTA CTG GG Mouse Hdac8 ACT ATT GC GGA GAT CCA ATG T CCT CCT AAA ATC AGA GTT GCC AG Mouse Hdac9 CAG AAG CAG CAC GAG AAT TTG A CTC TCT GCG ATG CCT CTC TAC Mouse Sik1 TCA TGT CGG AGT TCA GTG CG ACC TGC GTT TTG GTG ACT CG Mouse Sik2 CTG CTG GCA ACA TGG TGT G GGG AGA GTT GGT CCA TCA AAA G Mouse Sik3 GCC ATC CAC ACA TCA TCA GAC CCA AGT GGT CAA ATA TCT CCC C Mouse Lkb1 TTG GGC CTT TTC TCC GAG G CAG GTC CCC CAT CAG GTA CT Mouse Sun2 ATC CAG ACC TTC TAT TTC CAG GC CCC GGA AGC GGT AGA TAC AC Open in a separate window Primers used in this study table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Species Gene Probe Human ACTB Hs.PT.39a.22214847 Human RUNX2 Hs.PT.58.19252426 Human SPP1 Hs.PT.53a.19568141 Human HDAC4 Hs.PT.58.19252426 Open in a separate window Probes used in this study Cell transfection + siRNA Cells were transfected with pEGFP‐HDAC4, pEGFP‐HDAC4 3SA, pcDNA HDAC4‐Flag that were a gift from Tso‐Pang Yao (Addgene plasmid #45636, #45637, #30485) 59 or pEGFP‐HDAC4 3‐625 plasmid.

Techniques: Expressing, Binding Assay, Inhibition

Journal: EMBO Reports

Article Title: Salt‐inducible kinase induces cytoplasmic histone deacetylase 4 to promote vascular calcification

doi: 10.15252/embr.201643686

Figure Lengend Snippet: Probes used in this study

Article Snippet: All primers and probes were ordered from IDT (see Tables and ): table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Species Gene Forward Backward Human HDAC4 GGC CCA CCG GAA TCT GAA C GAA CTC TGG TCA AGG GAA CTG Mouse Actb CTC TGG CTC CTA GCA CCA TGA AGA GTA AAA CGC AGC TCA GTA ACA GTC CG Mouse Runx2 CGG CCC TCC CTG AAC TCT TGC CTG CCT GGG ATC TGT A Mouse Osteocalcin GCA ATA AGG TAG TGA ACA GAC TCC GTT TGT AGG CGG TCT TCA AGC Mouse Sox9 CGA CCC ATG AAC GCC TT GTC TCT TCT CGC TCT CGT TC Mouse Alkp GGC TGG AGA TGG ACA AAT TCC CCG AGT GGT AGT CAC AAT GCC Mouse Spp1 ATC TCA CCA TTC GGA TGA GTC T TGT AGG GAC GAT TGG AGT GAA A Mouse Enigma TGC AAG AAG AAG ATC ACT GGA G CAT TGA AGT CCT TGC CCC CT Mouse Hdac1 AGT CTG TTA CTA CTA CGA CGG G TGA GCA GCA AAT TGT GAG TCA T Mouse Hdac2 GGA GGA GGC TAC ACA ATC CG TCT GGA GTG TTC TGG TTT GTC A Mouse Hdac3 ACC GTG GCG TAT TTC TAC GAC CAG GCG ATG AGG TTT CAT TGG Mouse Hdac4 CAC TGC ATT TCC AGC GAT CC AAG ACG GGG TGG TTG TAG GA Mouse Hdac5 TGC AGC ACG TTT TGC TCC T GAC AGC TCC CCA GTT TTG GT Mouse Hdac6 TCC ACC GGC CAA GAT TCT TC CAG CAC ACT TCT TTC CAC CAC Mouse Hdac7 GAA CTC TTG AGC CCT TGG ACA GGT GTG CTG CTA CTA CTG GG Mouse Hdac8 ACT ATT GC GGA GAT CCA ATG T CCT CCT AAA ATC AGA GTT GCC AG Mouse Hdac9 CAG AAG CAG CAC GAG AAT TTG A CTC TCT GCG ATG CCT CTC TAC Mouse Sik1 TCA TGT CGG AGT TCA GTG CG ACC TGC GTT TTG GTG ACT CG Mouse Sik2 CTG CTG GCA ACA TGG TGT G GGG AGA GTT GGT CCA TCA AAA G Mouse Sik3 GCC ATC CAC ACA TCA TCA GAC CCA AGT GGT CAA ATA TCT CCC C Mouse Lkb1 TTG GGC CTT TTC TCC GAG G CAG GTC CCC CAT CAG GTA CT Mouse Sun2 ATC CAG ACC TTC TAT TTC CAG GC CCC GGA AGC GGT AGA TAC AC Open in a separate window Primers used in this study table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Species Gene Probe Human ACTB Hs.PT.39a.22214847 Human RUNX2 Hs.PT.58.19252426 Human SPP1 Hs.PT.53a.19568141 Human HDAC4 Hs.PT.58.19252426 Open in a separate window Probes used in this study Cell transfection + siRNA Cells were transfected with pEGFP‐HDAC4, pEGFP‐HDAC4 3SA, pcDNA HDAC4‐Flag that were a gift from Tso‐Pang Yao (Addgene plasmid #45636, #45637, #30485) 59 or pEGFP‐HDAC4 3‐625 plasmid.

Techniques:

Journal: EMBO Reports

Article Title: Salt‐inducible kinase induces cytoplasmic histone deacetylase 4 to promote vascular calcification

doi: 10.15252/embr.201643686

Figure Lengend Snippet: Primers used in this study

Article Snippet: All primers and probes were ordered from IDT (see Tables and ): table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Species Gene Forward Backward Human HDAC4 GGC CCA CCG GAA TCT GAA C GAA CTC TGG TCA AGG GAA CTG Mouse Actb CTC TGG CTC CTA GCA CCA TGA AGA GTA AAA CGC AGC TCA GTA ACA GTC CG Mouse Runx2 CGG CCC TCC CTG AAC TCT TGC CTG CCT GGG ATC TGT A Mouse Osteocalcin GCA ATA AGG TAG TGA ACA GAC TCC GTT TGT AGG CGG TCT TCA AGC Mouse Sox9 CGA CCC ATG AAC GCC TT GTC TCT TCT CGC TCT CGT TC Mouse Alkp GGC TGG AGA TGG ACA AAT TCC CCG AGT GGT AGT CAC AAT GCC Mouse Spp1 ATC TCA CCA TTC GGA TGA GTC T TGT AGG GAC GAT TGG AGT GAA A Mouse Enigma TGC AAG AAG AAG ATC ACT GGA G CAT TGA AGT CCT TGC CCC CT Mouse Hdac1 AGT CTG TTA CTA CTA CGA CGG G TGA GCA GCA AAT TGT GAG TCA T Mouse Hdac2 GGA GGA GGC TAC ACA ATC CG TCT GGA GTG TTC TGG TTT GTC A Mouse Hdac3 ACC GTG GCG TAT TTC TAC GAC CAG GCG ATG AGG TTT CAT TGG Mouse Hdac4 CAC TGC ATT TCC AGC GAT CC AAG ACG GGG TGG TTG TAG GA Mouse Hdac5 TGC AGC ACG TTT TGC TCC T GAC AGC TCC CCA GTT TTG GT Mouse Hdac6 TCC ACC GGC CAA GAT TCT TC CAG CAC ACT TCT TTC CAC CAC Mouse Hdac7 GAA CTC TTG AGC CCT TGG ACA GGT GTG CTG CTA CTA CTG GG Mouse Hdac8 ACT ATT GC GGA GAT CCA ATG T CCT CCT AAA ATC AGA GTT GCC AG Mouse Hdac9 CAG AAG CAG CAC GAG AAT TTG A CTC TCT GCG ATG CCT CTC TAC Mouse Sik1 TCA TGT CGG AGT TCA GTG CG ACC TGC GTT TTG GTG ACT CG Mouse Sik2 CTG CTG GCA ACA TGG TGT G GGG AGA GTT GGT CCA TCA AAA G Mouse Sik3 GCC ATC CAC ACA TCA TCA GAC CCA AGT GGT CAA ATA TCT CCC C Mouse Lkb1 TTG GGC CTT TTC TCC GAG G CAG GTC CCC CAT CAG GTA CT Mouse Sun2 ATC CAG ACC TTC TAT TTC CAG GC CCC GGA AGC GGT AGA TAC AC Open in a separate window Primers used in this study table ft1 table-wrap mode="anchored" t5 Table 2 caption a7 Species Gene Probe Human ACTB Hs.PT.39a.22214847 Human RUNX2 Hs.PT.58.19252426 Human SPP1 Hs.PT.53a.19568141 Human HDAC4 Hs.PT.58.19252426 Open in a separate window Probes used in this study Cell transfection + siRNA Cells were transfected with pEGFP‐HDAC4, pEGFP‐HDAC4 3SA, pcDNA HDAC4‐Flag that were a gift from Tso‐Pang Yao (Addgene plasmid #45636, #45637, #30485) 59 or pEGFP‐HDAC4 3‐625 plasmid.

Techniques: