Review



pcdna hdac4 flag  (Addgene inc)


Bioz Verified Symbol Addgene inc is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92

    Structured Review

    Addgene inc pcdna hdac4 flag
    Pcdna Hdac4 Flag, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4-FLAG+(Plasmid+%2330485)/pmc11730567-12-0-2
    Average 92 stars, based on 19 article reviews
    pcdna hdac4 flag - by Bioz Stars, 2026-09
    92/100 stars

    Images

    Related Articles

    other:

    Article Title: Protocol for in vitro observation of HDAC4 condensation during induced cardiac reprogramming
    Article Snippet: pcDNA-HDAC4-FLAG , Addgene , 30485.

    Transfection:

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

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

    Article Title: Salt-inducible kinase induces cytoplasmic histone deacetylase 4 to promote vascular calcification.
    Article Snippet: All primers and probes were ordered from IDT (see Tables 1 and 2): .. 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. ..

    Plasmid Preparation:

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

    Article Title: Novel role and regulation of HDAC4 in cocaine-related behaviors
    Article Snippet: The authors thank Dr. David Potter, Brandon Hughes, Yuhong Guo, Ben Zirlin and Emilia Pulver for technical assistance and Dr. Rachael Neve (Viral Core facility at MIT) for HSV-HDAC4 viruses. pcDNA-HDAC4.3SA-FLAG was a gift from Tso-Pang Yao (Addgene plasmid #30486). pcDNA-HDAC4-FLAG was a gift from Eric Verdin (Addgene plasmid #13821). .. The authors thank Dr. David Potter, Brandon Hughes, Yuhong Guo, Ben Zirlin and Emilia Pulver for technical assistance and Dr. Rachael Neve (Viral Core facility at MIT) for HSV-HDAC4 viruses. pcDNA-HDAC4.3SA-FLAG was a gift from Tso-Pang Yao (Addgene plasmid #30486). pcDNA-HDAC4-FLAG was a gift from Eric Verdin (Addgene plasmid #13821). ..

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

    Article Title: Salt-inducible kinase induces cytoplasmic histone deacetylase 4 to promote vascular calcification.
    Article Snippet: All primers and probes were ordered from IDT (see Tables 1 and 2): .. 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. ..

    Countercurrent Chromatography:

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



    Similar Products

    92
    Addgene inc pcdna hdac4 flag
    Pcdna Hdac4 Flag, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4-FLAG+(Plasmid+%2330485)/pmc11730567-12-0-2
    Average 92 stars, based on 1 article reviews
    pcdna hdac4 flag - by Bioz Stars, 2026-09
    92/100 stars
      Buy from Supplier

    93
    Addgene inc pcdna hdac4 3sa flag
    Pcdna Hdac4 3sa Flag, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4%2E3SA-FLAG+(Plasmid+%2330486)/pmc11730567-11-0-2
    Average 93 stars, based on 1 article reviews
    pcdna hdac4 3sa flag - by Bioz Stars, 2026-09
    93/100 stars
      Buy from Supplier

    92
    Addgene inc lab 3580 pcdna3 bad s112a s136a addgene
    Lab 3580 Pcdna3 Bad S112a S136a Addgene, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4-FLAG+(Plasmid+%2330485)/pmc11081035__NIHMS1988703___supplement___1-61-57-63
    Average 92 stars, based on 1 article reviews
    lab 3580 pcdna3 bad s112a s136a addgene - by Bioz Stars, 2026-09
    92/100 stars
      Buy from Supplier

    93
    Addgene inc lab pcdna hdac4 flag addgene 30485 tso pang yao
    Lab Pcdna Hdac4 Flag Addgene 30485 Tso Pang Yao, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4%2E3SA-FLAG+(Plasmid+%2330486)/pmc11081035__NIHMS1988703___supplement___1-61-51-53
    Average 93 stars, based on 1 article reviews
    lab pcdna hdac4 flag addgene 30485 tso pang yao - by Bioz Stars, 2026-09
    93/100 stars
      Buy from Supplier

    92
    Addgene inc hdac4 cdna
    ( a ) HDAC isoform gene expression in Cal27_VC and <t>Cal27_HDAC4.</t> Data shown are RPKM (reads per kilobase per million mapped reads) expression values normalized from three independent RNA sequencing experiments. The white bars represent the vector control cell line Cal27_VC, the black bars represent the HDAC4 overexpression clone Cal27_HDAC4. ( b ) Representative Western blot analysis of HDAC protein expression in vector control (VC), HDAC4 and HDAC5 overexpression clones of sensitive Cal27 and cisplatin resistant Cal27CisR cells. ( c ) Cisplatin concentration effect curves and IC 50 values for Cal27_VC and the corresponding cisplatin resistant subline Cal27CisR_VC measured by MTT assay. Resistance factor (RF) is 4-fold.
    Hdac4 Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4-FLAG+(Plasmid+%2330485)/pmc10056166-265-0-12
    Average 92 stars, based on 1 article reviews
    hdac4 cdna - by Bioz Stars, 2026-09
    92/100 stars
      Buy from Supplier

    92
    Addgene inc pcdna hdac4 flag plasmid
    ( a ) HDAC isoform gene expression in Cal27_VC and <t>Cal27_HDAC4.</t> Data shown are RPKM (reads per kilobase per million mapped reads) expression values normalized from three independent RNA sequencing experiments. The white bars represent the vector control cell line Cal27_VC, the black bars represent the HDAC4 overexpression clone Cal27_HDAC4. ( b ) Representative Western blot analysis of HDAC protein expression in vector control (VC), HDAC4 and HDAC5 overexpression clones of sensitive Cal27 and cisplatin resistant Cal27CisR cells. ( c ) Cisplatin concentration effect curves and IC 50 values for Cal27_VC and the corresponding cisplatin resistant subline Cal27CisR_VC measured by MTT assay. Resistance factor (RF) is 4-fold.
    Pcdna Hdac4 Flag Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4-FLAG+(Plasmid+%2330485)/pmc10056166-265-4-12
    Average 92 stars, based on 1 article reviews
    pcdna hdac4 flag plasmid - by Bioz Stars, 2026-09
    92/100 stars
      Buy from Supplier

    93
    Addgene inc hdac4 3sa
    TDPP degraded phospho-14-3-3-binding-motif embedded proteins. A. Western blot of phospho-(Ser)-14-3-3-binding-motif treated with control/TDPP vector with/without 10 nM OA. B. Illustration of the grey value distribution of phospho-(Ser)-14-3-3-binding-motif in each treatment in panel A. X axis represents the distance to the top edge of the blot results. Upper panel: lane 1 (black) vs. lane 3 (green). Lower panel: lane 2 (red) vs. lane 4 (blue). C. Quantification of phospho-(Ser)-14-3-3-binding-motifembedded proteins with different molecular weight in panel A. Different molecular weight ranges were grouped as different regions and marked as different colors, which were roughly grouped by similar band intensity and reflect the difference among treatments. D and E. Western blot results (D) and the quantifications (E) of several different phospho-14-3-3-binding-motif embedded proteins <t>(HDAC4,</t> HDAC5, FoxO1) in HEK293T/17 cells treated with control/TDPP vectors. F and G. Western blot result (F) and the quantification (G) of HDAC4 phospho-S632 in a HDAC4 wildtype vectors transfected HEK293T/17 cells treated with control/TDPP vector.
    Hdac4 3sa, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4%2E3SA-FLAG+(Plasmid+%2330486)/bio_rxiv__2022__11__22__517526-122-9-23
    Average 93 stars, based on 1 article reviews
    hdac4 3sa - by Bioz Stars, 2026-09
    93/100 stars
      Buy from Supplier

    92
    Addgene inc hdac4
    TDPP degraded phospho-14-3-3-binding-motif embedded proteins. A. Western blot of phospho-(Ser)-14-3-3-binding-motif treated with control/TDPP vector with/without 10 nM OA. B. Illustration of the grey value distribution of phospho-(Ser)-14-3-3-binding-motif in each treatment in panel A. X axis represents the distance to the top edge of the blot results. Upper panel: lane 1 (black) vs. lane 3 (green). Lower panel: lane 2 (red) vs. lane 4 (blue). C. Quantification of phospho-(Ser)-14-3-3-binding-motifembedded proteins with different molecular weight in panel A. Different molecular weight ranges were grouped as different regions and marked as different colors, which were roughly grouped by similar band intensity and reflect the difference among treatments. D and E. Western blot results (D) and the quantifications (E) of several different phospho-14-3-3-binding-motif embedded proteins <t>(HDAC4,</t> HDAC5, FoxO1) in HEK293T/17 cells treated with control/TDPP vectors. F and G. Western blot result (F) and the quantification (G) of HDAC4 phospho-S632 in a HDAC4 wildtype vectors transfected HEK293T/17 cells treated with control/TDPP vector.
    Hdac4, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4-FLAG+(Plasmid+%2330485)/bio_rxiv__2022__11__22__517526-122-7-23
    Average 92 stars, based on 1 article reviews
    hdac4 - by Bioz Stars, 2026-09
    92/100 stars
      Buy from Supplier

    92
    Addgene inc mammalian expression plasmids for hdac4
    <t>HDAC4</t> is decreased in H 2 O 2 - and UV-induced cellular senescence. ( A ) A scheme of H 2 O 2 - and UV-induced cellular senescence using human dermal fibroblasts ( B ) Senescence-associated beta-galactosidase assay was performed to detect senescent cells for control (left) and senescence-induced (right) cells (n = 4 each). Original magnification: x100. ( C ) HDAC4 expression analyzed by quantitative RT-PCR or western blotting. Alpha-tubulin was used as loading control. Data represent the mean ± SE (n = 4, ** P <0.01, *** P <0.001). C: control.
    Mammalian Expression Plasmids For Hdac4, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4-FLAG+(Plasmid+%2330485)/pmc09217707-126-1-14
    Average 92 stars, based on 1 article reviews
    mammalian expression plasmids for hdac4 - by Bioz Stars, 2026-09
    92/100 stars
      Buy from Supplier

    93
    Addgene inc hdac4 s a
    Figure 1 <t>HDAC4</t> expression and activity is modulated in the skeletal muscle of mdx and mdx;KO mice. (A) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in 6-week-old healthy controls (CTR), mdx, or mdx;KO GA muscles. Gapdh was used as loading control. The residual expression of HDAC4 in mdx;KO muscles is due to the presence of tissues other than muscle in the protein extract. Data are expressed as mean ± SEM, over CTR mice. n = 3 mice per condition. One-way ANOVA reveals a significant effect and interaction: *P < 0.05 mdx versus CTR and mdx;KO by Tukey’s HSD test. (B) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in CTR, mdx, and mdx;KO primary myotubes. Data are presented as mean ± SEM, over mdx cells. n = 3 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test. (C) WB analyses for HDAC4 in nuclear and cytoplasmic fractions of 6-week-old mdx skeletal muscles. Gapdh and H3 were used as loading control of cytoplasmic and nuclear fractions, respectively. (D) Representative IF for HDAC4 in primary mdx myotubes. Scale bar: 20 μm. (E) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in healthy and DMD patients’ muscles. Gapdh was used as loading control. Data are expressed as mean ± SEM, over healthy subjects. n = 3 mice per condition. (F) HDAC activity assay in CTR, mdx, and mdx;KO GA muscles at 6 weeks of age. Data are expressed as mean ± SEM, over CTR muscles. n = 6 mice per condition. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test. (G) Real-time PCR for class II HDAC members in 6-week-old CTR, mdx, and mdx;KO mice. Data are presented as mean ± SEM, over CTR muscles. n = 4/5 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test.
    Hdac4 S A, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pcdna+hdac4+flag/pcDNA-HDAC4%2E3SA-FLAG+(Plasmid+%2330486)/pm35170869-58-50-47
    Average 93 stars, based on 1 article reviews
    hdac4 s a - by Bioz Stars, 2026-09
    93/100 stars
      Buy from Supplier

    Image Search Results


    ( a ) HDAC isoform gene expression in Cal27_VC and Cal27_HDAC4. Data shown are RPKM (reads per kilobase per million mapped reads) expression values normalized from three independent RNA sequencing experiments. The white bars represent the vector control cell line Cal27_VC, the black bars represent the HDAC4 overexpression clone Cal27_HDAC4. ( b ) Representative Western blot analysis of HDAC protein expression in vector control (VC), HDAC4 and HDAC5 overexpression clones of sensitive Cal27 and cisplatin resistant Cal27CisR cells. ( c ) Cisplatin concentration effect curves and IC 50 values for Cal27_VC and the corresponding cisplatin resistant subline Cal27CisR_VC measured by MTT assay. Resistance factor (RF) is 4-fold.

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: ( a ) HDAC isoform gene expression in Cal27_VC and Cal27_HDAC4. Data shown are RPKM (reads per kilobase per million mapped reads) expression values normalized from three independent RNA sequencing experiments. The white bars represent the vector control cell line Cal27_VC, the black bars represent the HDAC4 overexpression clone Cal27_HDAC4. ( b ) Representative Western blot analysis of HDAC protein expression in vector control (VC), HDAC4 and HDAC5 overexpression clones of sensitive Cal27 and cisplatin resistant Cal27CisR cells. ( c ) Cisplatin concentration effect curves and IC 50 values for Cal27_VC and the corresponding cisplatin resistant subline Cal27CisR_VC measured by MTT assay. Resistance factor (RF) is 4-fold.

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Gene Expression, Expressing, RNA Sequencing, Plasmid Preparation, Control, Over Expression, Western Blot, Clone Assay, Concentration Assay, MTT Assay

    Effects of HDAC4 overexpression on cell proliferation analyzed over a time period up to 96 h by MTT assay. The value for 24 h was defined as a starting point with a relative proliferation set as 1. Values represent means ± SD of at least three independent experiments. Significance was calculated by t -test (* p ≤ 0.05, *** p ≤ 0.001).

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: Effects of HDAC4 overexpression on cell proliferation analyzed over a time period up to 96 h by MTT assay. The value for 24 h was defined as a starting point with a relative proliferation set as 1. Values represent means ± SD of at least three independent experiments. Significance was calculated by t -test (* p ≤ 0.05, *** p ≤ 0.001).

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Over Expression, MTT Assay

    Effects of HDAC4 overexpression and treatment with 5 µM CHDI0039 on tumor volume ( a ) and weight ( b ) in the chorioallantoic membrane (CAM) model. Tumors were seeded on the CAM and grown for 7 days. Treatment with CHDI0039 or buffer control was conducted at day 2 and day 4 after seeding. Values represent means ± SD of at least three independent experiments. Significance was calculated by t -test (ns: not significant; * p ≤ 0.05; ** p ≤ 0.01).

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: Effects of HDAC4 overexpression and treatment with 5 µM CHDI0039 on tumor volume ( a ) and weight ( b ) in the chorioallantoic membrane (CAM) model. Tumors were seeded on the CAM and grown for 7 days. Treatment with CHDI0039 or buffer control was conducted at day 2 and day 4 after seeding. Values represent means ± SD of at least three independent experiments. Significance was calculated by t -test (ns: not significant; * p ≤ 0.05; ** p ≤ 0.01).

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Over Expression, Membrane, Control

    Differentially expressed genes associated with  HDAC4  expression status and class IIa HDAC inhibition by 5 µM CHDI0039. Shown are the total number of differentially expressed genes (upper number) and the number of upregulated (up)- or downregulated (down) genes.

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: Differentially expressed genes associated with HDAC4 expression status and class IIa HDAC inhibition by 5 µM CHDI0039. Shown are the total number of differentially expressed genes (upper number) and the number of upregulated (up)- or downregulated (down) genes.

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Expressing, Inhibition

    Genes whose up- ( a ) or down- ( b ) regulation upon treatment with 5 µM CHDI0039 for 24 h in  Cal27_HDAC4  promote increased survival in head–neck cancer patients according to analysis by KM plotter database [ <xref ref-type= 32 ]." width="100%" height="100%">

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: Genes whose up- ( a ) or down- ( b ) regulation upon treatment with 5 µM CHDI0039 for 24 h in Cal27_HDAC4 promote increased survival in head–neck cancer patients according to analysis by KM plotter database [ 32 ].

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques:

    Cytotoxic activity (MTT assay) of vorinostat, CHDI0039, bortezomib, and cisplatin at Cal27 and Cal27CisR vector control (VC) and  HDAC4  and HDAC5 overexpression clones after compound incubation for 72 h. Shown are IC 50 values ± SD of at least three independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: Cytotoxic activity (MTT assay) of vorinostat, CHDI0039, bortezomib, and cisplatin at Cal27 and Cal27CisR vector control (VC) and HDAC4 and HDAC5 overexpression clones after compound incubation for 72 h. Shown are IC 50 values ± SD of at least three independent experiments.

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Activity Assay, MTT Assay, Plasmid Preparation, Control, Over Expression, Clone Assay, Incubation

    IC 50 values of cisplatin [µM] in the absence and presence of 2.5 μM/5 μM CHDI0039 or 0.75 µM vorinostat. ns (not significant), *** ( p ≤ 0.001). SD ≤ 10% of the mean.

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: IC 50 values of cisplatin [µM] in the absence and presence of 2.5 μM/5 μM CHDI0039 or 0.75 µM vorinostat. ns (not significant), *** ( p ≤ 0.001). SD ≤ 10% of the mean.

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Control

    Coincubation of CHDI0039 with bortezomib sensitizes Cal27CisR cells with overexpression of  HDAC4,  HDAC5, or vector control (VC) to bortezomib. Shown are IC 50 values of bortezomib [nM] and shift factors [SF]. † = *** ( p ≤ 0.001). SD ≤ 15% of the mean. Values represent means of three independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: Coincubation of CHDI0039 with bortezomib sensitizes Cal27CisR cells with overexpression of HDAC4, HDAC5, or vector control (VC) to bortezomib. Shown are IC 50 values of bortezomib [nM] and shift factors [SF]. † = *** ( p ≤ 0.001). SD ≤ 15% of the mean. Values represent means of three independent experiments.

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Over Expression, Plasmid Preparation, Control

    CI values according to Chou–Talalay of the drug combination CHDI0039 and bortezomib in Cal27CisR_VC,  Cal27CisR_HDAC4,  and Cal27CisR_HDAC5. * = fraction affected less than 20%. Values represent means of at least three independent experiments.

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: CI values according to Chou–Talalay of the drug combination CHDI0039 and bortezomib in Cal27CisR_VC, Cal27CisR_HDAC4, and Cal27CisR_HDAC5. * = fraction affected less than 20%. Values represent means of at least three independent experiments.

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques:

    Representative fluorescent images of various Cal27CisR cell clones either untreated (control) or treated with 10 µM CHDI0039, or 10 nM bortezomib, or the combination thereof for 24 h. Hoechst 33342 (“Hoechst”, blue color) was used as nuclear staining and CellEvent Caspase-3/7 green detection reagent (green color) was used for caspase 3/7 activation. ( a ) Cal27CisR_VC cells. ( b ) Cal27CisR_HDAC4 cells. ( c ) Cal27CisR_HDAC5 cells. ( d ) Caspase 3/7 activation by single treatment and combinations of different concentrations of CHDI0039 (CHDI) and 10 nM bortezomib (Bort) in Cal27CisR_VC, Cal27CisR_HDAC4, and Cal27CisR_HDAC5. Cells were incubated with CHDI0039, bortezomib, or a combination thereof for 24 h. An amount of 0.5 µM Staurosporin was incubated for 8 h and served as positive control. Caspase 3/7 activation was analyzed by ArrayScan XTI. Data are the mean ± SD of two experiments, each with three replicates. Statistical analysis to compare the caspase 3/7 activation of the indicated treatments was performed using t -test. ns p > 0.05, * ( p ≤ 0.05), ** ( p ≤ 0.01), *** ( p ≤ 0.001), **** ( p ≤ 0.0001).

    Journal: International Journal of Molecular Sciences

    Article Title: Synergistic Interaction of the Class IIa HDAC Inhibitor CHDI0039 with Bortezomib in Head and Neck Cancer Cells

    doi: 10.3390/ijms24065553

    Figure Lengend Snippet: Representative fluorescent images of various Cal27CisR cell clones either untreated (control) or treated with 10 µM CHDI0039, or 10 nM bortezomib, or the combination thereof for 24 h. Hoechst 33342 (“Hoechst”, blue color) was used as nuclear staining and CellEvent Caspase-3/7 green detection reagent (green color) was used for caspase 3/7 activation. ( a ) Cal27CisR_VC cells. ( b ) Cal27CisR_HDAC4 cells. ( c ) Cal27CisR_HDAC5 cells. ( d ) Caspase 3/7 activation by single treatment and combinations of different concentrations of CHDI0039 (CHDI) and 10 nM bortezomib (Bort) in Cal27CisR_VC, Cal27CisR_HDAC4, and Cal27CisR_HDAC5. Cells were incubated with CHDI0039, bortezomib, or a combination thereof for 24 h. An amount of 0.5 µM Staurosporin was incubated for 8 h and served as positive control. Caspase 3/7 activation was analyzed by ArrayScan XTI. Data are the mean ± SD of two experiments, each with three replicates. Statistical analysis to compare the caspase 3/7 activation of the indicated treatments was performed using t -test. ns p > 0.05, * ( p ≤ 0.05), ** ( p ≤ 0.01), *** ( p ≤ 0.001), **** ( p ≤ 0.0001).

    Article Snippet: HDAC4 cDNA from the pcDNA-HDAC4-FLAG plasmid was kindly provided by Tso-Pang Yao (Addgene, Watertown, MA, USA, plasmid #30485).

    Techniques: Clone Assay, Control, Staining, Activation Assay, Incubation, Positive Control

    TDPP degraded phospho-14-3-3-binding-motif embedded proteins. A. Western blot of phospho-(Ser)-14-3-3-binding-motif treated with control/TDPP vector with/without 10 nM OA. B. Illustration of the grey value distribution of phospho-(Ser)-14-3-3-binding-motif in each treatment in panel A. X axis represents the distance to the top edge of the blot results. Upper panel: lane 1 (black) vs. lane 3 (green). Lower panel: lane 2 (red) vs. lane 4 (blue). C. Quantification of phospho-(Ser)-14-3-3-binding-motifembedded proteins with different molecular weight in panel A. Different molecular weight ranges were grouped as different regions and marked as different colors, which were roughly grouped by similar band intensity and reflect the difference among treatments. D and E. Western blot results (D) and the quantifications (E) of several different phospho-14-3-3-binding-motif embedded proteins (HDAC4, HDAC5, FoxO1) in HEK293T/17 cells treated with control/TDPP vectors. F and G. Western blot result (F) and the quantification (G) of HDAC4 phospho-S632 in a HDAC4 wildtype vectors transfected HEK293T/17 cells treated with control/TDPP vector.

    Journal: bioRxiv

    Article Title: A ubiquitination-mediated degradation system to target phospho-14-3-3-binding-motif embedded proteins

    doi: 10.1101/2022.11.22.517526

    Figure Lengend Snippet: TDPP degraded phospho-14-3-3-binding-motif embedded proteins. A. Western blot of phospho-(Ser)-14-3-3-binding-motif treated with control/TDPP vector with/without 10 nM OA. B. Illustration of the grey value distribution of phospho-(Ser)-14-3-3-binding-motif in each treatment in panel A. X axis represents the distance to the top edge of the blot results. Upper panel: lane 1 (black) vs. lane 3 (green). Lower panel: lane 2 (red) vs. lane 4 (blue). C. Quantification of phospho-(Ser)-14-3-3-binding-motifembedded proteins with different molecular weight in panel A. Different molecular weight ranges were grouped as different regions and marked as different colors, which were roughly grouped by similar band intensity and reflect the difference among treatments. D and E. Western blot results (D) and the quantifications (E) of several different phospho-14-3-3-binding-motif embedded proteins (HDAC4, HDAC5, FoxO1) in HEK293T/17 cells treated with control/TDPP vectors. F and G. Western blot result (F) and the quantification (G) of HDAC4 phospho-S632 in a HDAC4 wildtype vectors transfected HEK293T/17 cells treated with control/TDPP vector.

    Article Snippet: Plasmids encoded with VHL (#19234), 14-3-3ζ (#1942), HDAC4 (#30485), HDAC4 3SA (#30486), HDAC5 (#32213), FoxO1 (#9022), CFL1 (#78295), BICRA (#34902) were obtained from Addgene.

    Techniques: Binding Assay, Western Blot, Control, Plasmid Preparation, Molecular Weight, Transfection

    TDPP degraded phospho-14-3-3-binding-motif embedded proteins. A. Western blot of phospho-(Ser)-14-3-3-binding-motif treated with control/TDPP vector with/without 10 nM OA. B. Illustration of the grey value distribution of phospho-(Ser)-14-3-3-binding-motif in each treatment in panel A. X axis represents the distance to the top edge of the blot results. Upper panel: lane 1 (black) vs. lane 3 (green). Lower panel: lane 2 (red) vs. lane 4 (blue). C. Quantification of phospho-(Ser)-14-3-3-binding-motifembedded proteins with different molecular weight in panel A. Different molecular weight ranges were grouped as different regions and marked as different colors, which were roughly grouped by similar band intensity and reflect the difference among treatments. D and E. Western blot results (D) and the quantifications (E) of several different phospho-14-3-3-binding-motif embedded proteins (HDAC4, HDAC5, FoxO1) in HEK293T/17 cells treated with control/TDPP vectors. F and G. Western blot result (F) and the quantification (G) of HDAC4 phospho-S632 in a HDAC4 wildtype vectors transfected HEK293T/17 cells treated with control/TDPP vector.

    Journal: bioRxiv

    Article Title: A ubiquitination-mediated degradation system to target phospho-14-3-3-binding-motif embedded proteins

    doi: 10.1101/2022.11.22.517526

    Figure Lengend Snippet: TDPP degraded phospho-14-3-3-binding-motif embedded proteins. A. Western blot of phospho-(Ser)-14-3-3-binding-motif treated with control/TDPP vector with/without 10 nM OA. B. Illustration of the grey value distribution of phospho-(Ser)-14-3-3-binding-motif in each treatment in panel A. X axis represents the distance to the top edge of the blot results. Upper panel: lane 1 (black) vs. lane 3 (green). Lower panel: lane 2 (red) vs. lane 4 (blue). C. Quantification of phospho-(Ser)-14-3-3-binding-motifembedded proteins with different molecular weight in panel A. Different molecular weight ranges were grouped as different regions and marked as different colors, which were roughly grouped by similar band intensity and reflect the difference among treatments. D and E. Western blot results (D) and the quantifications (E) of several different phospho-14-3-3-binding-motif embedded proteins (HDAC4, HDAC5, FoxO1) in HEK293T/17 cells treated with control/TDPP vectors. F and G. Western blot result (F) and the quantification (G) of HDAC4 phospho-S632 in a HDAC4 wildtype vectors transfected HEK293T/17 cells treated with control/TDPP vector.

    Article Snippet: Plasmids encoded with VHL (#19234), 14-3-3ζ (#1942), HDAC4 (#30485), HDAC4 3SA (#30486), HDAC5 (#32213), FoxO1 (#9022), CFL1 (#78295), BICRA (#34902) were obtained from Addgene.

    Techniques: Binding Assay, Western Blot, Control, Plasmid Preparation, Molecular Weight, Transfection

    HDAC4 is decreased in H 2 O 2 - and UV-induced cellular senescence. ( A ) A scheme of H 2 O 2 - and UV-induced cellular senescence using human dermal fibroblasts ( B ) Senescence-associated beta-galactosidase assay was performed to detect senescent cells for control (left) and senescence-induced (right) cells (n = 4 each). Original magnification: x100. ( C ) HDAC4 expression analyzed by quantitative RT-PCR or western blotting. Alpha-tubulin was used as loading control. Data represent the mean ± SE (n = 4, ** P <0.01, *** P <0.001). C: control.

    Journal: Aging (Albany NY)

    Article Title: Histone deacetylase 4 reverses cellular senescence via DDIT4 in dermal fibroblasts

    doi: 10.18632/aging.204118

    Figure Lengend Snippet: HDAC4 is decreased in H 2 O 2 - and UV-induced cellular senescence. ( A ) A scheme of H 2 O 2 - and UV-induced cellular senescence using human dermal fibroblasts ( B ) Senescence-associated beta-galactosidase assay was performed to detect senescent cells for control (left) and senescence-induced (right) cells (n = 4 each). Original magnification: x100. ( C ) HDAC4 expression analyzed by quantitative RT-PCR or western blotting. Alpha-tubulin was used as loading control. Data represent the mean ± SE (n = 4, ** P <0.01, *** P <0.001). C: control.

    Article Snippet: The mammalian expression plasmids for HDAC4 (plasmid #30485) and DDIT4 (RC202847) were purchased from Addgene (Cambridge, MA, USA) and OriGene (Rockville, MD, USA), respectively.

    Techniques: β-Gal Assay, Control, Expressing, Quantitative RT-PCR, Western Blot

    Integrative transcriptome analysis identifies DDIT4 as a candidate target. ( A ) Venn diagram showing the relationships among DEGs identified from the three comparisons: HDAC4 overexpression versus knockdown (HDAC4 targets); UV-induced senescent versus control HDFs (UV-induced senescence); and H 2 O 2 -induced senescent versus control HDFs (H 2 O 2 -induced senescence). Numbers in parentheses, numbers of DEGs. ( B ) Differential expression patterns of the five overlapping DEGs in the three comparisons (see legend). ( C ) The mRNA levels of DDIT4, GINS2, and PGAP6 analyzed by RT-PCR (n=5). Data represent the mean ± SE (n = 5, * P < 0.05, ** P <0.01, *** P <0.001). NC: scrambled control.

    Journal: Aging (Albany NY)

    Article Title: Histone deacetylase 4 reverses cellular senescence via DDIT4 in dermal fibroblasts

    doi: 10.18632/aging.204118

    Figure Lengend Snippet: Integrative transcriptome analysis identifies DDIT4 as a candidate target. ( A ) Venn diagram showing the relationships among DEGs identified from the three comparisons: HDAC4 overexpression versus knockdown (HDAC4 targets); UV-induced senescent versus control HDFs (UV-induced senescence); and H 2 O 2 -induced senescent versus control HDFs (H 2 O 2 -induced senescence). Numbers in parentheses, numbers of DEGs. ( B ) Differential expression patterns of the five overlapping DEGs in the three comparisons (see legend). ( C ) The mRNA levels of DDIT4, GINS2, and PGAP6 analyzed by RT-PCR (n=5). Data represent the mean ± SE (n = 5, * P < 0.05, ** P <0.01, *** P <0.001). NC: scrambled control.

    Article Snippet: The mammalian expression plasmids for HDAC4 (plasmid #30485) and DDIT4 (RC202847) were purchased from Addgene (Cambridge, MA, USA) and OriGene (Rockville, MD, USA), respectively.

    Techniques: Over Expression, Knockdown, Control, Quantitative Proteomics, Reverse Transcription Polymerase Chain Reaction

    Senescence-induced reduction of DDIT4 is restored by HDAC4 overexpression. ( A ) Human dermal fibroblasts were transfected with negative control scrambled siRNA (NC) or HDAC4 siRNA. ( B ) For overexpression of HDAC4, cells were treated with pcDNA3 or HDAC4 plasmid DNA and the medium was replaced after 6 h. ( A – D ) RT-PCR and western blotting were performed to determine the expression of DDIT4 and HDAC4. Data are presented as mean ± SE (n = 5, * P <0.05, ** P <0.01, *** P <0.001 vs. NC, pcDNA3, or Control). ( E , F ) After senescence was induced, cells were transfected with pcDNA3 or HDAC4 overexpression vector. DDIT4, p21, and HDAC4 protein expression were analyzed by western blotting. Beta-actin and alpha-tubulin served as a loading control. Data are presented as mean ± SE using Student’s t-test (n = 5, * P <0.05, ** P <0.01, *** P <0.001 control vs H 2 O 2 ; # P <0.05, ## P <0.01, pcDNA3 vs HDAC4). C: control.

    Journal: Aging (Albany NY)

    Article Title: Histone deacetylase 4 reverses cellular senescence via DDIT4 in dermal fibroblasts

    doi: 10.18632/aging.204118

    Figure Lengend Snippet: Senescence-induced reduction of DDIT4 is restored by HDAC4 overexpression. ( A ) Human dermal fibroblasts were transfected with negative control scrambled siRNA (NC) or HDAC4 siRNA. ( B ) For overexpression of HDAC4, cells were treated with pcDNA3 or HDAC4 plasmid DNA and the medium was replaced after 6 h. ( A – D ) RT-PCR and western blotting were performed to determine the expression of DDIT4 and HDAC4. Data are presented as mean ± SE (n = 5, * P <0.05, ** P <0.01, *** P <0.001 vs. NC, pcDNA3, or Control). ( E , F ) After senescence was induced, cells were transfected with pcDNA3 or HDAC4 overexpression vector. DDIT4, p21, and HDAC4 protein expression were analyzed by western blotting. Beta-actin and alpha-tubulin served as a loading control. Data are presented as mean ± SE using Student’s t-test (n = 5, * P <0.05, ** P <0.01, *** P <0.001 control vs H 2 O 2 ; # P <0.05, ## P <0.01, pcDNA3 vs HDAC4). C: control.

    Article Snippet: The mammalian expression plasmids for HDAC4 (plasmid #30485) and DDIT4 (RC202847) were purchased from Addgene (Cambridge, MA, USA) and OriGene (Rockville, MD, USA), respectively.

    Techniques: Over Expression, Transfection, Negative Control, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Control

    Inhibition of DDIT4 exacerbates cellular senescence. ( A , B ) Primary dermal fibroblasts were subject to senescence induction with H 2 O 2 or UV. After senescence induction, cells were transfected with negative siRNA or DDIT4 siRNA for 48 h. Beta-galactosidase assay was performed to detect senescent cells. Original magnification: x100. Data represent the mean ± SE (n=3, * P <0.05, ** P <0.01 vs siDDIT4; # P <0.05, ## P <0.01, pCDH vs HDAC4). C: control, NC: scrambled control.

    Journal: Aging (Albany NY)

    Article Title: Histone deacetylase 4 reverses cellular senescence via DDIT4 in dermal fibroblasts

    doi: 10.18632/aging.204118

    Figure Lengend Snippet: Inhibition of DDIT4 exacerbates cellular senescence. ( A , B ) Primary dermal fibroblasts were subject to senescence induction with H 2 O 2 or UV. After senescence induction, cells were transfected with negative siRNA or DDIT4 siRNA for 48 h. Beta-galactosidase assay was performed to detect senescent cells. Original magnification: x100. Data represent the mean ± SE (n=3, * P <0.05, ** P <0.01 vs siDDIT4; # P <0.05, ## P <0.01, pCDH vs HDAC4). C: control, NC: scrambled control.

    Article Snippet: The mammalian expression plasmids for HDAC4 (plasmid #30485) and DDIT4 (RC202847) were purchased from Addgene (Cambridge, MA, USA) and OriGene (Rockville, MD, USA), respectively.

    Techniques: Inhibition, Transfection, β-Gal Assay, Control

    Figure 1 HDAC4 expression and activity is modulated in the skeletal muscle of mdx and mdx;KO mice. (A) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in 6-week-old healthy controls (CTR), mdx, or mdx;KO GA muscles. Gapdh was used as loading control. The residual expression of HDAC4 in mdx;KO muscles is due to the presence of tissues other than muscle in the protein extract. Data are expressed as mean ± SEM, over CTR mice. n = 3 mice per condition. One-way ANOVA reveals a significant effect and interaction: *P < 0.05 mdx versus CTR and mdx;KO by Tukey’s HSD test. (B) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in CTR, mdx, and mdx;KO primary myotubes. Data are presented as mean ± SEM, over mdx cells. n = 3 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test. (C) WB analyses for HDAC4 in nuclear and cytoplasmic fractions of 6-week-old mdx skeletal muscles. Gapdh and H3 were used as loading control of cytoplasmic and nuclear fractions, respectively. (D) Representative IF for HDAC4 in primary mdx myotubes. Scale bar: 20 μm. (E) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in healthy and DMD patients’ muscles. Gapdh was used as loading control. Data are expressed as mean ± SEM, over healthy subjects. n = 3 mice per condition. (F) HDAC activity assay in CTR, mdx, and mdx;KO GA muscles at 6 weeks of age. Data are expressed as mean ± SEM, over CTR muscles. n = 6 mice per condition. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test. (G) Real-time PCR for class II HDAC members in 6-week-old CTR, mdx, and mdx;KO mice. Data are presented as mean ± SEM, over CTR muscles. n = 4/5 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test.

    Journal: Journal of cachexia, sarcopenia and muscle

    Article Title: Cytoplasmic HDAC4 regulates the membrane repair mechanism in Duchenne muscular dystrophy.

    doi: 10.1002/jcsm.12891

    Figure Lengend Snippet: Figure 1 HDAC4 expression and activity is modulated in the skeletal muscle of mdx and mdx;KO mice. (A) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in 6-week-old healthy controls (CTR), mdx, or mdx;KO GA muscles. Gapdh was used as loading control. The residual expression of HDAC4 in mdx;KO muscles is due to the presence of tissues other than muscle in the protein extract. Data are expressed as mean ± SEM, over CTR mice. n = 3 mice per condition. One-way ANOVA reveals a significant effect and interaction: *P < 0.05 mdx versus CTR and mdx;KO by Tukey’s HSD test. (B) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in CTR, mdx, and mdx;KO primary myotubes. Data are presented as mean ± SEM, over mdx cells. n = 3 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test. (C) WB analyses for HDAC4 in nuclear and cytoplasmic fractions of 6-week-old mdx skeletal muscles. Gapdh and H3 were used as loading control of cytoplasmic and nuclear fractions, respectively. (D) Representative IF for HDAC4 in primary mdx myotubes. Scale bar: 20 μm. (E) Densitometric analysis of the WB bands for HDAC4, over Gapdh, in healthy and DMD patients’ muscles. Gapdh was used as loading control. Data are expressed as mean ± SEM, over healthy subjects. n = 3 mice per condition. (F) HDAC activity assay in CTR, mdx, and mdx;KO GA muscles at 6 weeks of age. Data are expressed as mean ± SEM, over CTR muscles. n = 6 mice per condition. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test. (G) Real-time PCR for class II HDAC members in 6-week-old CTR, mdx, and mdx;KO mice. Data are presented as mean ± SEM, over CTR muscles. n = 4/5 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test.

    Article Snippet: The following plasmids were used: pCDNA3-N2myc (Stratagene); Snap-GFP (kindly provided by Tullio Pozzan); GFP-HDAC4 (generously provided by Eric N. Olson); hHDAC4 [NM_006037.3]*L175A/Myc (CliniSciences) or GFP-hHDAC4 [NM_006037.3]*L175A (generated by Claudio Brancolini’s laboratory) (named HDAC4 L/A); GFP-hHDAC4 [NM_006037.3]*L175A D840N (called GFP-HDAC4 L/A D840N) (generated by Claudio Brancolini’s laboratory); pcDNA-HDAC4.3SA-FLAG (Addgene, #30486) (named HDAC4 S/A); EGFP-Dysferlin (kindly provided by Simone Spuler); and Trim72 (Myc-DDK-tagged) (named c-myc-Trim72) (OriGene).

    Techniques: Expressing, Activity Assay, Muscles, Control, HDAC Activity Assay, Real-time Polymerase Chain Reaction

    Figure 2 The deletion of HDAC4 in skeletal muscle exacerbates the pathological features of muscular dystrophy in mice. (A) Representative pictures of mdx and mdx;KO GA muscle labelled with IgG at 3 and 6 weeks of age. Scale bar: 500 μm. Quantification of the IgG-positive fibre CSA, over muscle CSA, of mdx and mdx;KO GA muscles at 3 and 6 weeks of age. Data are expressed as mean ± SEM. n = 3 mice per genotype, at each time point. *P < 0.05; **P < 0.005 by Student’s t-test. (B) Creatine kinase levels in sera of mdx and mdx;KO mice at 6 weeks of age. Data are expressed as mean ± SEM, over mdx mice. n = 7 mice per genotype. *P < 0.05 by Student’s t-test. (C) Densitometric analyses of the WB bands for Rip3 protein in mdx and mdx;KO GA muscles, at 6 weeks of age. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 3 mice for each genotype. #P < 0.01 by Student’s t-test. (D) Densitometric analyses of the WB bands for Myogenin protein in GA muscles of 6-week-old mdx and mdx; KO mice. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 4/5 mice for each genotype. #P < 0.01 by Student’s t-test. (E) Representative IF for Myh3 (red) and laminin (green) in GA muscles of mdx and mdx;KO mice at 6 weeks of age and quantification of the Myh3-positive CSA. Scale bar: 100 μm. Data are expressed as mean ± SEM. n = 3 mice per genotype. **P < 0.005 by Student’s t-test. (F) Muscle performance of mdx and mdx;KO mice by treadmill test, at 6 weeks, 6 months, and 16 months. Data are presented as mean ± SEM. n = 6 mice per genotype. *P < 0.05 by Student’s t-test.

    Journal: Journal of cachexia, sarcopenia and muscle

    Article Title: Cytoplasmic HDAC4 regulates the membrane repair mechanism in Duchenne muscular dystrophy.

    doi: 10.1002/jcsm.12891

    Figure Lengend Snippet: Figure 2 The deletion of HDAC4 in skeletal muscle exacerbates the pathological features of muscular dystrophy in mice. (A) Representative pictures of mdx and mdx;KO GA muscle labelled with IgG at 3 and 6 weeks of age. Scale bar: 500 μm. Quantification of the IgG-positive fibre CSA, over muscle CSA, of mdx and mdx;KO GA muscles at 3 and 6 weeks of age. Data are expressed as mean ± SEM. n = 3 mice per genotype, at each time point. *P < 0.05; **P < 0.005 by Student’s t-test. (B) Creatine kinase levels in sera of mdx and mdx;KO mice at 6 weeks of age. Data are expressed as mean ± SEM, over mdx mice. n = 7 mice per genotype. *P < 0.05 by Student’s t-test. (C) Densitometric analyses of the WB bands for Rip3 protein in mdx and mdx;KO GA muscles, at 6 weeks of age. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 3 mice for each genotype. #P < 0.01 by Student’s t-test. (D) Densitometric analyses of the WB bands for Myogenin protein in GA muscles of 6-week-old mdx and mdx; KO mice. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 4/5 mice for each genotype. #P < 0.01 by Student’s t-test. (E) Representative IF for Myh3 (red) and laminin (green) in GA muscles of mdx and mdx;KO mice at 6 weeks of age and quantification of the Myh3-positive CSA. Scale bar: 100 μm. Data are expressed as mean ± SEM. n = 3 mice per genotype. **P < 0.005 by Student’s t-test. (F) Muscle performance of mdx and mdx;KO mice by treadmill test, at 6 weeks, 6 months, and 16 months. Data are presented as mean ± SEM. n = 6 mice per genotype. *P < 0.05 by Student’s t-test.

    Article Snippet: The following plasmids were used: pCDNA3-N2myc (Stratagene); Snap-GFP (kindly provided by Tullio Pozzan); GFP-HDAC4 (generously provided by Eric N. Olson); hHDAC4 [NM_006037.3]*L175A/Myc (CliniSciences) or GFP-hHDAC4 [NM_006037.3]*L175A (generated by Claudio Brancolini’s laboratory) (named HDAC4 L/A); GFP-hHDAC4 [NM_006037.3]*L175A D840N (called GFP-HDAC4 L/A D840N) (generated by Claudio Brancolini’s laboratory); pcDNA-HDAC4.3SA-FLAG (Addgene, #30486) (named HDAC4 S/A); EGFP-Dysferlin (kindly provided by Simone Spuler); and Trim72 (Myc-DDK-tagged) (named c-myc-Trim72) (OriGene).

    Techniques: Muscles, Staining, Control

    Figure 4 HDAC4 affects membrane stability and is necessary for an adequate expression and localization of proteins involved in the membrane re- pair mechanism in mdx mice. (A) Representative images of mdx and mdx;KO GA muscle at 6 months of age with or without treadmill exercise, la- belled with EBD and quantification of EBD-positive fibre CSA, over muscle CSA. Scale bars: 500 and 100 μm. Data are expressed as mean ± SEM. n = 5 mice per genotype. Two-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD post hoc test. (B) Den- sitometric analyses of the WB bands for Dysferlin and Trim72 proteins in CTR, mdx, and mdx;KO GA muscles, at 6 weeks of age. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 3–6 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; **P < 0.005 by Tukey’s HSD post hoc test. (C) Representative images of CTR, mdx, and mdx;KO GA muscle at 6 weeks of age, labelled with Dysferlin (red) or Trim72 (green). Scale bar: 100 μm. (D) Densitometric analyses of the WB bands for Dysferlin and Trim72 proteins in CTR, mdx, and mdx;KO primary muscle cells. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 3–6 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: #P < 0.01 by Tukey’s HSD post hoc test. (E) Representative images of CTR, mdx, and mdx;KO differentiated primary muscle cells labelled with Dysferlin (green) or Trim72 (red). Scale bar: 50 μm.

    Journal: Journal of cachexia, sarcopenia and muscle

    Article Title: Cytoplasmic HDAC4 regulates the membrane repair mechanism in Duchenne muscular dystrophy.

    doi: 10.1002/jcsm.12891

    Figure Lengend Snippet: Figure 4 HDAC4 affects membrane stability and is necessary for an adequate expression and localization of proteins involved in the membrane re- pair mechanism in mdx mice. (A) Representative images of mdx and mdx;KO GA muscle at 6 months of age with or without treadmill exercise, la- belled with EBD and quantification of EBD-positive fibre CSA, over muscle CSA. Scale bars: 500 and 100 μm. Data are expressed as mean ± SEM. n = 5 mice per genotype. Two-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD post hoc test. (B) Den- sitometric analyses of the WB bands for Dysferlin and Trim72 proteins in CTR, mdx, and mdx;KO GA muscles, at 6 weeks of age. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 3–6 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; **P < 0.005 by Tukey’s HSD post hoc test. (C) Representative images of CTR, mdx, and mdx;KO GA muscle at 6 weeks of age, labelled with Dysferlin (red) or Trim72 (green). Scale bar: 100 μm. (D) Densitometric analyses of the WB bands for Dysferlin and Trim72 proteins in CTR, mdx, and mdx;KO primary muscle cells. Stain-free protein bands were used as loading control. Data are shown as mean ± SEM, over mdx mice. n = 3–6 mice for each genotype. One-way ANOVA reveals a significant effect and interaction: #P < 0.01 by Tukey’s HSD post hoc test. (E) Representative images of CTR, mdx, and mdx;KO differentiated primary muscle cells labelled with Dysferlin (green) or Trim72 (red). Scale bar: 50 μm.

    Article Snippet: The following plasmids were used: pCDNA3-N2myc (Stratagene); Snap-GFP (kindly provided by Tullio Pozzan); GFP-HDAC4 (generously provided by Eric N. Olson); hHDAC4 [NM_006037.3]*L175A/Myc (CliniSciences) or GFP-hHDAC4 [NM_006037.3]*L175A (generated by Claudio Brancolini’s laboratory) (named HDAC4 L/A); GFP-hHDAC4 [NM_006037.3]*L175A D840N (called GFP-HDAC4 L/A D840N) (generated by Claudio Brancolini’s laboratory); pcDNA-HDAC4.3SA-FLAG (Addgene, #30486) (named HDAC4 S/A); EGFP-Dysferlin (kindly provided by Simone Spuler); and Trim72 (Myc-DDK-tagged) (named c-myc-Trim72) (OriGene).

    Techniques: Membrane, Expressing, Muscles, Staining, Control

    Figure 6 The ectopic expression of cytoplasmic-restricted HDAC4 ameliorates mdx;KO phenotype in vitro and in vivo. (A) Representative IF for GFP in mdx;KO primary muscle cells transfected with GFP, GFP-HDAC4, HDAC4 S/A + GFP, or GFP-HDAC4 L/A after 3 days of differentiation. Scale bar: 50 μm. HDAC4 expression in mdx;KO muscle cells transfected with GFP-HDAC4 or HDAC4 S/A + GFP or GFP-HDAC4 L/A expressing plasmid, over GFP- transfected ones, by real-time PCR. Data are presented as mean ± SEM. n = 3 mdx;KO mice; **P < 0.005; #P < 0.01 versus GFP-transfected cells by Student’s t-test. Quantification of fusion index and number of muscle cells. n = 3 mdx;KO mice. Data are presented as mean ± SEM. *P < 0.05; #P < 0.01 versus GFP-transfected cells by Student’s t-test. (B) Representative images of mdx;KO GA muscles electroporated with either HDAC4 L/A expressing vector or GFP, as control, stained with H&E. Scale bars: 500 and 100 μm. (C) HDAC4 expression in mdx;KO GA muscles electroporated with either HDAC4 L/A expressing plasmid or GFP, as control, by WB analysis. Stain-free protein bands were used as a loading control. Data are shown as mean ± SEM, over GFP-electroporated muscles. n = 4/5 muscles per condition. #P < 0.01 by Student’s t-test. (D) Quantification of EBD-positive fibre CSA, over muscle CSA, in electroporated mdx;KO muscles. Data are expressed as mean ± SEM. n = 4/5 mice per condition. **P < 0.005 by Student’s t- test. (E) Densitometric analyses of the WB bands for Rip3 protein in mdx;KO GA electroporated muscles. Stain-free protein bands were used as a load- ing control. Data are expressed as mean ± SEM, over GFP-electroporated muscles. n = 4 mice per condition. *P < 0.05 by Student’s t-test. (F) Densi- tometric analyses of the WB bands for Myogenin and Myh3 protein levels in mdx;KO GA electroporated muscles. Stain-free protein bands were used as a loading control. Data are shown as mean ± SEM, over GFP-electroporated muscles. n = 4/5 muscles per condition. *P < 0.05; #P < 0.01 by Student’s t-test. (G) Densitometric analyses of the WB bands for Dysferlin and Trim72 protein levels in mdx;KO GA electroporated muscles. Stain-free protein bands were used as a loading control. Data are shown as mean ± SEM, over GFP-electroporated muscles. n = 4 muscles per condition. *P < 0.05; #P < 0.01 by Student’s t-test. (H) Muscle performance of mdx;KO mice electroporated with either HDAC4 L/A expressing vector or GFP, as control, by treadmill test. Data are presented as mean ± SEM. n = 4/5 mice per condition. *P < 0.05 by Student’s t-test.

    Journal: Journal of cachexia, sarcopenia and muscle

    Article Title: Cytoplasmic HDAC4 regulates the membrane repair mechanism in Duchenne muscular dystrophy.

    doi: 10.1002/jcsm.12891

    Figure Lengend Snippet: Figure 6 The ectopic expression of cytoplasmic-restricted HDAC4 ameliorates mdx;KO phenotype in vitro and in vivo. (A) Representative IF for GFP in mdx;KO primary muscle cells transfected with GFP, GFP-HDAC4, HDAC4 S/A + GFP, or GFP-HDAC4 L/A after 3 days of differentiation. Scale bar: 50 μm. HDAC4 expression in mdx;KO muscle cells transfected with GFP-HDAC4 or HDAC4 S/A + GFP or GFP-HDAC4 L/A expressing plasmid, over GFP- transfected ones, by real-time PCR. Data are presented as mean ± SEM. n = 3 mdx;KO mice; **P < 0.005; #P < 0.01 versus GFP-transfected cells by Student’s t-test. Quantification of fusion index and number of muscle cells. n = 3 mdx;KO mice. Data are presented as mean ± SEM. *P < 0.05; #P < 0.01 versus GFP-transfected cells by Student’s t-test. (B) Representative images of mdx;KO GA muscles electroporated with either HDAC4 L/A expressing vector or GFP, as control, stained with H&E. Scale bars: 500 and 100 μm. (C) HDAC4 expression in mdx;KO GA muscles electroporated with either HDAC4 L/A expressing plasmid or GFP, as control, by WB analysis. Stain-free protein bands were used as a loading control. Data are shown as mean ± SEM, over GFP-electroporated muscles. n = 4/5 muscles per condition. #P < 0.01 by Student’s t-test. (D) Quantification of EBD-positive fibre CSA, over muscle CSA, in electroporated mdx;KO muscles. Data are expressed as mean ± SEM. n = 4/5 mice per condition. **P < 0.005 by Student’s t- test. (E) Densitometric analyses of the WB bands for Rip3 protein in mdx;KO GA electroporated muscles. Stain-free protein bands were used as a load- ing control. Data are expressed as mean ± SEM, over GFP-electroporated muscles. n = 4 mice per condition. *P < 0.05 by Student’s t-test. (F) Densi- tometric analyses of the WB bands for Myogenin and Myh3 protein levels in mdx;KO GA electroporated muscles. Stain-free protein bands were used as a loading control. Data are shown as mean ± SEM, over GFP-electroporated muscles. n = 4/5 muscles per condition. *P < 0.05; #P < 0.01 by Student’s t-test. (G) Densitometric analyses of the WB bands for Dysferlin and Trim72 protein levels in mdx;KO GA electroporated muscles. Stain-free protein bands were used as a loading control. Data are shown as mean ± SEM, over GFP-electroporated muscles. n = 4 muscles per condition. *P < 0.05; #P < 0.01 by Student’s t-test. (H) Muscle performance of mdx;KO mice electroporated with either HDAC4 L/A expressing vector or GFP, as control, by treadmill test. Data are presented as mean ± SEM. n = 4/5 mice per condition. *P < 0.05 by Student’s t-test.

    Article Snippet: The following plasmids were used: pCDNA3-N2myc (Stratagene); Snap-GFP (kindly provided by Tullio Pozzan); GFP-HDAC4 (generously provided by Eric N. Olson); hHDAC4 [NM_006037.3]*L175A/Myc (CliniSciences) or GFP-hHDAC4 [NM_006037.3]*L175A (generated by Claudio Brancolini’s laboratory) (named HDAC4 L/A); GFP-hHDAC4 [NM_006037.3]*L175A D840N (called GFP-HDAC4 L/A D840N) (generated by Claudio Brancolini’s laboratory); pcDNA-HDAC4.3SA-FLAG (Addgene, #30486) (named HDAC4 S/A); EGFP-Dysferlin (kindly provided by Simone Spuler); and Trim72 (Myc-DDK-tagged) (named c-myc-Trim72) (OriGene).

    Techniques: Expressing, In Vitro, In Vivo, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Muscles, Control, Staining

    Figure 7 HDAC4 modulates Trim72 mRNA stability in mdx satellite cells. (A) Representative IF for GFP in mdx;KO primary muscle cells transfected with GFP-HDAC4, GFP-HDAC4 L/A, GFP-HDAC4 L/A D840N, or GFP-expressing plasmids and terminally differentiated. Scale bar: 50 μm. Quantification of the fusion index and number of muscle cells. Data are presented as mean ± SEM. n = 3 mdx;KO mice. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD post hoc test. (B) Real-time PCR analyses of transcript levels of Trim72 in mdx (light blue) and mdx;KO (dark blue) myotubes treated with Actinomycin D. The relative expression of Trim72 at different time points was calculated by normalizing the gene expression in Actinomycin D-treated mdx or mdx;KO samples over untreated samples (t = 0). Data are presented as mean ± SEM. n = 3. One-way ANOVA reveals a significant effect: #P < 0.01 mdx or mdx;KO-treated samples versus t = 0 by Tukey’s HSD test. (C) Expression levels of Trim72 in mdx;KO transfected muscle cells with GFP or GFP-HDAC4 L/A, over mdx SCs, by real-time PCR. Data are presented as mean ± SEM. n = 3 mice per genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test.

    Journal: Journal of cachexia, sarcopenia and muscle

    Article Title: Cytoplasmic HDAC4 regulates the membrane repair mechanism in Duchenne muscular dystrophy.

    doi: 10.1002/jcsm.12891

    Figure Lengend Snippet: Figure 7 HDAC4 modulates Trim72 mRNA stability in mdx satellite cells. (A) Representative IF for GFP in mdx;KO primary muscle cells transfected with GFP-HDAC4, GFP-HDAC4 L/A, GFP-HDAC4 L/A D840N, or GFP-expressing plasmids and terminally differentiated. Scale bar: 50 μm. Quantification of the fusion index and number of muscle cells. Data are presented as mean ± SEM. n = 3 mdx;KO mice. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD post hoc test. (B) Real-time PCR analyses of transcript levels of Trim72 in mdx (light blue) and mdx;KO (dark blue) myotubes treated with Actinomycin D. The relative expression of Trim72 at different time points was calculated by normalizing the gene expression in Actinomycin D-treated mdx or mdx;KO samples over untreated samples (t = 0). Data are presented as mean ± SEM. n = 3. One-way ANOVA reveals a significant effect: #P < 0.01 mdx or mdx;KO-treated samples versus t = 0 by Tukey’s HSD test. (C) Expression levels of Trim72 in mdx;KO transfected muscle cells with GFP or GFP-HDAC4 L/A, over mdx SCs, by real-time PCR. Data are presented as mean ± SEM. n = 3 mice per genotype. One-way ANOVA reveals a significant effect and interaction: *P < 0.05; #P < 0.01 by Tukey’s HSD test.

    Article Snippet: The following plasmids were used: pCDNA3-N2myc (Stratagene); Snap-GFP (kindly provided by Tullio Pozzan); GFP-HDAC4 (generously provided by Eric N. Olson); hHDAC4 [NM_006037.3]*L175A/Myc (CliniSciences) or GFP-hHDAC4 [NM_006037.3]*L175A (generated by Claudio Brancolini’s laboratory) (named HDAC4 L/A); GFP-hHDAC4 [NM_006037.3]*L175A D840N (called GFP-HDAC4 L/A D840N) (generated by Claudio Brancolini’s laboratory); pcDNA-HDAC4.3SA-FLAG (Addgene, #30486) (named HDAC4 S/A); EGFP-Dysferlin (kindly provided by Simone Spuler); and Trim72 (Myc-DDK-tagged) (named c-myc-Trim72) (OriGene).

    Techniques: Transfection, Expressing, Real-time Polymerase Chain Reaction, Gene Expression

    Figure 8 Model of the proposed role of HDAC4 in DMD muscles. Cytoplasmic HDAC4 mediates the membrane repair mechanism response, thus pro- moting muscle stem cell survival and fusion in vitro, restoring muscle integrity, and favouring de novo myogenesis in vivo. Taken together, these func- tions result in improved muscle architecture and function in mdx mice.

    Journal: Journal of cachexia, sarcopenia and muscle

    Article Title: Cytoplasmic HDAC4 regulates the membrane repair mechanism in Duchenne muscular dystrophy.

    doi: 10.1002/jcsm.12891

    Figure Lengend Snippet: Figure 8 Model of the proposed role of HDAC4 in DMD muscles. Cytoplasmic HDAC4 mediates the membrane repair mechanism response, thus pro- moting muscle stem cell survival and fusion in vitro, restoring muscle integrity, and favouring de novo myogenesis in vivo. Taken together, these func- tions result in improved muscle architecture and function in mdx mice.

    Article Snippet: The following plasmids were used: pCDNA3-N2myc (Stratagene); Snap-GFP (kindly provided by Tullio Pozzan); GFP-HDAC4 (generously provided by Eric N. Olson); hHDAC4 [NM_006037.3]*L175A/Myc (CliniSciences) or GFP-hHDAC4 [NM_006037.3]*L175A (generated by Claudio Brancolini’s laboratory) (named HDAC4 L/A); GFP-hHDAC4 [NM_006037.3]*L175A D840N (called GFP-HDAC4 L/A D840N) (generated by Claudio Brancolini’s laboratory); pcDNA-HDAC4.3SA-FLAG (Addgene, #30486) (named HDAC4 S/A); EGFP-Dysferlin (kindly provided by Simone Spuler); and Trim72 (Myc-DDK-tagged) (named c-myc-Trim72) (OriGene).

    Techniques: Muscles, Membrane, In Vitro, In Vivo