hdac4 inhibitor lmk 235  (MedChemExpress)

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the NAT10-HDAC4 positive feedback loop counteracts immunosuppression in breast cancer

doi: 10.1186/s13046-025-03638-7

Figure Lengend Snippet: NAT10 stimulates HDAC4 expression via ac4C modification. A Volcano plot of differentially expressed ac4C acetylation peaks ( P < 0.05) identified by acRIP-seq in NAT10 knockdown cells. B Distribution of ac4C peaks across mRNA regions in breast cancer cells. C Sequence logo of the enriched motif within ac4C peaks identified by HOMER. D Volcano plot of differentially expressed mRNAs ( P < 0.05) identified by RNA-seq in NAT10 knockdown cells. E Integrative analysis of acRIP-seq and RNA-seq data to identify potential downstream targets of NAT10. F , G Relative HDAC4 mRNA expression measured by qRT-PCR after NAT10 knockdown. H , I acRIP-qPCR analysis of ac4C modification ( H ) and RIP-qPCR analysis of NAT10 binding on HDAC4 mRNA ( I ) after NAT10 knockdown. J Genome browser view of ac4C peaks on HDAC4 mRNA from acRIP-seq. K Relative HDAC4 protein levels measured by Western blot after NAT10 knockdown. L qRT-PCR analysis of HDAC4 mRNA stability after actinomycin D treatment in NAT10 knockdown cells. M Luciferase activity of the reporter constructs containing the wild-type or ac4C site mutated sequence in NAT10 knockdown cells. N , O HDAC4 expression assessed by qRT-PCR ( N ) and Western blot ( O ) after transfection with oeNAT10 or NAT10 G641E mutant plasmid. P qRT-PCR analysis of HDAC4 mRNA stability after actinomycin D treatment in cells transfected with oeNAT10 or NAT10 G641E . Q , R acRIP-qPCR analysis of ac4C modification ( Q ) and RIP-qPCR analysis of NAT10 binding on HDAC4 mRNA ( R ) after transfection with oeNAT10 or NAT10 G641E . S Luciferase activity of the reporter constructs containing the wild-type or ac4C site mutated sequence after transfection with oeNAT10 or NAT10 G641E . All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The HDAC4 inhibitor LMK-235 (MedChemExpress, #HY18998, China) was administered intraperitoneally at doses of 5 mg/kg or 10 mg/kg body weight every three days.

Techniques: Expressing, Modification, Knockdown, Sequencing, RNA Sequencing, Quantitative RT-PCR, Binding Assay, Western Blot, Luciferase, Activity Assay, Construct, Transfection, Mutagenesis, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the NAT10-HDAC4 positive feedback loop counteracts immunosuppression in breast cancer

doi: 10.1186/s13046-025-03638-7

Figure Lengend Snippet: HDAC4 acts as a deacetylase to regulate NAT10 stability. A Representative IHC images and quantification of NAT10 and HDAC4 expression in breast cancer tissues ( n = 220; scale bar, 200 µm). B , C Correlation between NAT10 and HDAC4 expression levels in breast cancer tissues, as analyzed by IHC ( B ) and in the TCGA-BRCA RNA-seq dataset ( C) . D , E Relative NAT10 expression was detected by Western blot ( D ) and qRT-PCR ( E ) after HDAC4 knockdown or overexpression. F Western blot analysis of NAT10 protein stability after cycloheximide treatment in cells with HDAC4 knockdown or overexpression. G Molecular docking model of the NAT10 (blue) and HDAC4 (yellow) interaction, with an enlarged view highlighting predicted hydrogen bonds. H Co-IP followed by Western blot analysis was used to detect the interaction between endogenous HDAC4 and NAT10. I Co-IP followed by Western blot analysis assessed NAT10 acetylation levels after HDAC4 knockdown or overexpression. J Prediction of potential deacetylation sites on NAT10 using MusiteDeep. K The predicted acetylation sites on NAT10 were individually mutated (K→R), and the effects of HDAC4 on the acetylation levels of the six NAT10 mutants were examined in 293T cells. L Detection of NAT10 acetylation in cells expressing NAT10 K354R following transfection with vector or HDAC4. M Detection of NAT10 acetylation in cells transfected with wild-type or catalytically inactive (D840N) HDAC4. N Western blot analysis of NAT10 protein stability following cycloheximide treatment in cells transfected with siNC, siHDAC4, or siHDAC4 plus NAT10 K354R . All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The HDAC4 inhibitor LMK-235 (MedChemExpress, #HY18998, China) was administered intraperitoneally at doses of 5 mg/kg or 10 mg/kg body weight every three days.

Techniques: Histone Deacetylase Assay, Expressing, RNA Sequencing, Western Blot, Quantitative RT-PCR, Knockdown, Over Expression, Co-Immunoprecipitation Assay, Transfection, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the NAT10-HDAC4 positive feedback loop counteracts immunosuppression in breast cancer

doi: 10.1186/s13046-025-03638-7

Figure Lengend Snippet: The proliferative function of NAT10 in breast cancer depends on HDAC4. A CCK-8 assays of cells transfected with vector, oeNAT10, oeNAT10 plus siHDAC4-1, or oeNAT10 plus siHDAC4-2. B Flow cytometry analysis of apoptotic rates (LR + UR) in cells transfected with the indicated constructs. C Flow cytometry analysis of cell cycle distribution in cells transfected with the indicated constructs. D Representative images of tumors in mice treated with vector, oeNAT10, oeNAT10 plus LMK235 (5 mg/kg), or oeNAT10 plus LMK235 (10 mg/kg) ( n = 6). E Tumor growth curves and tumor weight analysis in tumors from different groups ( n = 6). F Representative IHC images of NAT10, HDAC4, and Ki67 in tumor tissues from different treatment groups, with quantification of staining intensity ( n = 6; scale bars, 50 μm). G Representative TUNEL staining of tumor tissues from different treatment groups, with quantification of staining intensity ( n = 6; scale bars, 100 px). H-J CCK-8 assays ( H ) and flow cytometry analyses of apoptotic rates ( I ) and cell cycle distribution ( J ) in cells transfected with vector, oeNAT10, oeNAT10 plus LMK235 (1 µM), or oeNAT10 plus LMK235 (2 µM). All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The HDAC4 inhibitor LMK-235 (MedChemExpress, #HY18998, China) was administered intraperitoneally at doses of 5 mg/kg or 10 mg/kg body weight every three days.

Techniques: CCK-8 Assay, Transfection, Plasmid Preparation, Flow Cytometry, Construct, Staining, TUNEL Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the NAT10-HDAC4 positive feedback loop counteracts immunosuppression in breast cancer

doi: 10.1186/s13046-025-03638-7

Figure Lengend Snippet: NAT10 promotes PD-L1 expression through HDAC4. A Relative PD-L1 expression assessed by Western blot and qRT–PCR following NAT10 knockdown or overexpression. B IF staining of PD-L1 in cells with NAT10 knockdown or overexpression (scale bars, 5 μm). C Representative IHC images of PD-L1 in tumor tissues from shNC or shNAT10 groups and vector or oeNAT10 groups, with quantification of staining intensity ( n = 6; scale bars, 50 μm). D-F Relative PD-L1 expression analyzed by Western blot ( D ), qRT-PCR ( D ), and IF ( E-F ) in cells transfected with vector, oeNAT10, oeNAT10 plus siHDAC4-1, or oeNAT10 plus siHDAC4-2. G Representative IHC images of PD-L1 in tumor tissues from vector, oeNAT10, oeNAT10 plus LMK235 (5 mg/kg), or oeNAT10 plus LMK235 (10 mg/kg), with quantification of staining intensity ( n = 6; scale bars, 50 μm) All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The HDAC4 inhibitor LMK-235 (MedChemExpress, #HY18998, China) was administered intraperitoneally at doses of 5 mg/kg or 10 mg/kg body weight every three days.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Knockdown, Over Expression, Staining, Plasmid Preparation, Transfection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the NAT10-HDAC4 positive feedback loop counteracts immunosuppression in breast cancer

doi: 10.1186/s13046-025-03638-7

Figure Lengend Snippet: NAT10 promotes PD-L1 expression through the HDAC4–NF-κB pathway. A GSEA of RNA-seq (siHDAC4 vs. siNC) data showed enrichment of the NF-κB signaling pathway (NES, normalized enrichment score; P value by permutation test). B Correlation analysis of p65, NAT10, HDAC4, and PD-L1 mRNA expression in the TCGA RNA-seq dataset. C Relative PD-L1 expression assessed by qRT-PCR following HDAC4 knockdown or overexpression. D Western blot analysis of PD-L1 and NF-κB pathway proteins upon HDAC4 knockdown or overexpression. E Predicted p65 binding sites in the PD-L1 promoter identified using the JASPAR database. F ChIP-qPCR analysis of p65 enrichment at the PD-L1 promoter after HDAC4 knockdown or overexpression. G Western blot analysis of NF-κB pathway proteins in cells transfected with vector, oeNAT10, oeNAT10 plus siHDAC4-1, or oeNAT10 plus siHDAC4-2. All data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: The HDAC4 inhibitor LMK-235 (MedChemExpress, #HY18998, China) was administered intraperitoneally at doses of 5 mg/kg or 10 mg/kg body weight every three days.

Techniques: Expressing, RNA Sequencing, Quantitative RT-PCR, Knockdown, Over Expression, Western Blot, Binding Assay, ChIP-qPCR, Transfection, Plasmid Preparation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Targeting the NAT10-HDAC4 positive feedback loop counteracts immunosuppression in breast cancer

doi: 10.1186/s13046-025-03638-7

Figure Lengend Snippet: Mode pattern of the NAT10/HDAC4/NF-κB regulatory network in breast cancer. NAT10 mediated ac4C modification stabilizes HDAC4 mRNA, while HDAC4 stabilizes NAT10 protein, forming a reciprocal regulatory loop. HDAC4 activates NF-κB signaling, leading to PD-L1 upregulation and immune evasion. Inhibition of the NAT10/HDAC4/NF-κB axis reduces PD-L1 expression and restores antitumor immunity in breast cancer

Article Snippet: The HDAC4 inhibitor LMK-235 (MedChemExpress, #HY18998, China) was administered intraperitoneally at doses of 5 mg/kg or 10 mg/kg body weight every three days.

Techniques: Modification, Inhibition, Expressing

Journal: Environmental pollution (Barking, Essex : 1987)

Article Title: Silica's silent threat: Contributing to skin fibrosis in systemic sclerosis by targeting the HDAC4/Smad2/3 pathway.

doi: 10.1016/j.envpol.2024.124194

Figure Lengend Snippet: Fig. 5. HDAC4 promoted the development of silica-induced skin fibrosis by increasing the expression of collagen and pro-fibrotic factors via the Smad2/3 pathway. (A–C) Relative mRNA and protein expression levels of HDAC4, COL1, α-SMA, and CTGF in normal HSFs transfected with OE-HDAC4 using qRT-PCR and western blotting. (D–F) Relative mRNA and protein expression levels of HDAC4, COL1, α-SMA, and CTGF in SSc HSFs transfected with si-HDAC4 using qRT-PCR and western blotting. (G–H) Expression of phosphorylated Smad2/3 proteins in normal HSFs transfected with OE-HDAC4 or SSc HSFs transfected with si-HDAC4 using western blotting. (I) Cell proliferation activity of HSFs treated with different doses of LMK235 for different durations using CCK-8 (n = 6). (J–L) Relative mRNA and protein expression levels of HDAC4, COL1, α-SMA, CTGF, and phosphorylated Smad2/3 in SSc HSFs that were stimulated with silica nanopowders (SiO2) and then treated with LMK235 using qRT-PCR and western blotting. Data are presented as mean ± standard deviation (n = 3); statistical analysis by Student’s t-test or ANOVA: *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 versus the indicated group.

Article Snippet: The mice in the SiO2 and SiO2+BLM groups were subdivided into SiO2-Ctrl, SiO2-LMK235, SiO2+BLM-Ctrl, and SiO2+BLM-LMK235 groups (n = 5), and given daily intraperitoneal injections of the HDAC4-selective inhibitor LMK235 (20 mg/kg (#7569, Selleck, USA)) (Fan et al., 2021; Wang et al., 2019) or LMK235-free drug solvent (Ctrl) as appropriate for 4 weeks.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Western Blot, Activity Assay, CCK-8 Assay, Standard Deviation

Journal: Environmental pollution (Barking, Essex : 1987)

Article Title: Silica's silent threat: Contributing to skin fibrosis in systemic sclerosis by targeting the HDAC4/Smad2/3 pathway.

doi: 10.1016/j.envpol.2024.124194

Figure Lengend Snippet: Fig. 6. HDAC4 inhibition attenuated silica-induced collagen deposition and skin fibrosis in mice. The mice in the SiO2 and SiO2+BLM groups were subdivided into SiO2-Ctrl, SiO2-LMK235, SiO2+BLM-Ctrl, and SiO2+BLM-LMK235 groups, and given daily intraperitoneal injections of LMK235 (20 mg/kg) or LMK235-free drug solvent (Ctrl) as appropriate for 4 weeks. (A) HE and Masson staining of the dorsal skin lesions of the differentially stimulated mice to visualize the extent of dermatopathological alterations and fibrosis (100 × ). The arrows indicate the dermal thickness. (B) Dermal thickness of the dorsal skin lesions of the differentially stimulated mice. (C–G) qRT-PCR and western blotting analysis of the relative mRNA and protein expression levels of HDAC4, COL1, α-SMA, and CTGF in the dorsal skin lesions of the differentially stimulated mice. Data are presented as mean ± standard deviation (n = 5); statistical analysis by ANOVA: **P < 0.01, ***P < 0.001, and ****P < 0.0001 versus the indicated group.

Article Snippet: The mice in the SiO2 and SiO2+BLM groups were subdivided into SiO2-Ctrl, SiO2-LMK235, SiO2+BLM-Ctrl, and SiO2+BLM-LMK235 groups (n = 5), and given daily intraperitoneal injections of the HDAC4-selective inhibitor LMK235 (20 mg/kg (#7569, Selleck, USA)) (Fan et al., 2021; Wang et al., 2019) or LMK235-free drug solvent (Ctrl) as appropriate for 4 weeks.

Techniques: Inhibition, Solvent, Staining, Quantitative RT-PCR, Western Blot, Expressing, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: HDAC4 induces the development of asthma by increasing Slug-upregulated CXCL12 expression through KLF5 deacetylation

doi: 10.1186/s12967-021-02812-7

Figure Lengend Snippet: Inhibition of HDAC4 attenuates airway inflammation and remodeling in asthmatic mice. a The silencing efficiency of HDAC4 as determined by RT-qPCR. b The silencing efficiency of HDAC4 as determined by Western blot assay. c The expression of HDAC4 in lung tissues of mice of mice with different treatment as determined by RT-qPCR. d The expression of HDAC4 in lung tissues of mice with different treatment as determined by Western blot assay. e The AHR in mice with different treatment as examined by methacholine challenge test. f The number of inflammatory cells in BALF of mice with different treatment. g The expression of IL-4, IL-5, and IL-13 in BALF of mice with different treatment as determined by ELISA. h The infiltration of inflammatory cells around the pulmonary bronchus of mice with different treatment as examined by HE staining (200 × ; 50 μm). i The expression of α-SMA determined to measure the area of BSMCs in the lung tissues as examined by immunohistochemistry. j The expression of hydroxyproline determined by Western blot assay in the lung tissues of mice with different treatment determined to examine the content of collagen in the lung tissues. * p < 0.05 vs. sh-NC. # p < 0.05 vs. sham-operated mice. These data were measurement data, expressed as mean ± standard deviation. Data between two groups were compared using unpaired t -test. Data among multiple groups were compared using one-way ANOVA, combined with Tukey’s post-hoc tests. Repeated measures ANOVA was performed for comparison on data at different time points among each group, combined with Bonferroni post-hoc tests. n = 6

Article Snippet: LMK-235, an HDAC4 deacetylase inhibitor (2 nM, HY-18998, MCE), further suppressed the proliferation and migration of BSMCs.

Techniques: Inhibition, Quantitative RT-PCR, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemistry, Standard Deviation, Comparison

Journal: Journal of Translational Medicine

Article Title: HDAC4 induces the development of asthma by increasing Slug-upregulated CXCL12 expression through KLF5 deacetylation

doi: 10.1186/s12967-021-02812-7

Figure Lengend Snippet: HDAC4 promotes airway inflammation and remodeling by increasing KLF5 transcriptional activity through deacetylation. a The expression of HDAC4 in BEAS-2B and HBE cells with different treatment as determined by RT-qPCR. b The acetylation level of HDAC4 and KLF5 as determined by Western blot assay. c The binding between HDAC4 and KLF5 as examined by IP assay. d The transcription activity of KLF5 as examined by dual luciferase reporter gene assay. e The expression of TGF-β and inflammatory factors IL-4, IL-5, and IL-13 in the supernatant of BEAS-2B and HBE cells with different treatment as examined by ELISA. f The proliferation of BSMCs with different treatment as examined by CCK-8 assay. g The proliferation of BSMCs with different treatments as examined by Transwell. * p < 0.05 vs. control or control-CM. # p < 0.05 vs. OVA + sh-NC. & p < 0.05 vs. OVA + sh-HDAC4 + vector. These data are measurement data, expressed as mean ± standard deviation. Data among multiple groups were compared using one-way ANOVA, combined with Tukey’s post-hoc tests

Article Snippet: LMK-235, an HDAC4 deacetylase inhibitor (2 nM, HY-18998, MCE), further suppressed the proliferation and migration of BSMCs.

Techniques: Activity Assay, Expressing, Quantitative RT-PCR, Western Blot, Binding Assay, Luciferase, Reporter Gene Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Control, Plasmid Preparation, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: HDAC4 induces the development of asthma by increasing Slug-upregulated CXCL12 expression through KLF5 deacetylation

doi: 10.1186/s12967-021-02812-7

Figure Lengend Snippet: HDAC4 promotes the proliferation and migration of BSMCs via regulation of the KLF5/Slug/CXCL12 axis. a The expression of HDAC4, KLF5, Slug and CXCL12 as determined by RT-qPCR. b The expression of HDAC4, KLF5, Ac-KLF5, Slug, and CXCL12 as determined by Western blot assay. c The expression of TGF-β and inflammatory factors IL-4, IL-5, and IL-13 in supernatant of BEAS-2B and HBE cells with different treatment as examined by ELISA. d The proliferation of BSMCs with different treatments as examined by CCK-8 assay. e The migration of BSMCs with different treatments as examined by Transwell assay. * p < 0.05 vs. sh-NC or control. # p < 0.05 vs. OVA + sh-NC. & vs. OVA + sh-HDAC4 + vector. These data are measurement data, expressed as mean ± standard deviation. Data among multiple groups were compared using one-way ANOVA, combined with Tukey’s post-hoc tests

Article Snippet: LMK-235, an HDAC4 deacetylase inhibitor (2 nM, HY-18998, MCE), further suppressed the proliferation and migration of BSMCs.

Techniques: Migration, Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Transwell Assay, Control, Plasmid Preparation, Standard Deviation

Journal: Journal of Translational Medicine

Article Title: HDAC4 induces the development of asthma by increasing Slug-upregulated CXCL12 expression through KLF5 deacetylation

doi: 10.1186/s12967-021-02812-7

Figure Lengend Snippet: HDAC4 promotes airway inflammation and remodeling in asthmatic mice via regulation of the KLF5/Slug/CXCL12 axis. a The expression of HDAC4, KLF5, Slug, and CXCL12 in the lung tissues of mice with different treatments as determined by RT-qPCR. b The expression of HDAC4, KLF5, Ac-KLF5, Slug, and CXCL12 in the lung tissues of mice with different treatments. c The AHR in mice with different treatments as examined by methacholine challenge test. d The number of inflammatory cells in BALF of mice with different treatment. e The expression of IL-4, IL-5, and IL-13 in BALF of mice with different treatments as determined by ELISA. f The expression of α-SMA determined to measure the area of BSMCs in the lung tissues as examined by immunohistochemistry. * p < 0.05 vs. sh-NC. # p < 0.05 vs. sham-operated mice. These data were measurement data, expressed as mean ± standard deviation. Data among multiple groups were compared using one-way ANOVA, combined with Tukey’s post-hoc tests. Repeated measures ANOVA was performed for comparison on data at different time points among each group, combined with Bonferroni post-hoc tests. n = 6

Article Snippet: LMK-235, an HDAC4 deacetylase inhibitor (2 nM, HY-18998, MCE), further suppressed the proliferation and migration of BSMCs.

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Standard Deviation, Comparison

Journal: Journal of Translational Medicine

Article Title: HDAC4 induces the development of asthma by increasing Slug-upregulated CXCL12 expression through KLF5 deacetylation

doi: 10.1186/s12967-021-02812-7

Figure Lengend Snippet: Molecular mechanism regarding HDAC4-mediated KLF5/Slug/CXCL12 axis in asthma. HDAC4 deacetylates KLF5 and increases its transcriptional activity. The deacetylated KLF5 promotes the expression of Slug in the promoter region of Slug, and then promotes the expression of CXCL12. This causes inflammation in bronchial epithelial cells and then induces the proliferation and migration of BSMCs, which leads to airway remodeling and thus aggravates the progression of asthma in mice

Article Snippet: LMK-235, an HDAC4 deacetylase inhibitor (2 nM, HY-18998, MCE), further suppressed the proliferation and migration of BSMCs.

Techniques: Activity Assay, Expressing, Migration

Journal: Frontiers in Cellular Neuroscience

Article Title: Inhibition of HDAC4 Attenuated JNK/c-Jun-Dependent Neuronal Apoptosis and Early Brain Injury Following Subarachnoid Hemorrhage by Transcriptionally Suppressing MKK7

doi: 10.3389/fncel.2019.00468

Figure Lengend Snippet: Inhibition of HDAC4 transcriptionally downregulated MKK7 expression and lowered 5K-evoked JNK/c-Jun activity. (A) Human glioma cell line U251 was transfected with siNC or the small interference RNAs (siRNAs) against indicated HDAC members , and MKK7 mRNA levels were detected by Q-PCR. Mean ± SD, * P < 0.05 vs. siNC. (B) Rat C6 glioma cells were transfected with siNC, siHDAC4-a, or siHDAC4-b, and mkk7 mRNA or MKK7 protein levels were determined by Q-PCR or WB, respectively. Mean ± SD, * P < 0.05 vs. siNC. (C) DIV5 CGNs transfected with siNC, siHDAC4-a, or siHDAC4-b together with pGFP plasmids were subjected to IF for HDAC4 and MKK7 co-staining. Photos were obtained using a confocal microscope (scale bar = 20 μm). GFP was used to mark the transfected cells, and the effect of HDAC4 knockdown on MKK7 expression was determined by scoring the percentage of the GFP-positive neuron population with HDAC4 or MKK7 stained. Mean ± SD, * P < 0.05 vs. siNC, # P < 0.05 vs. siNC. (D–F) CGNs in 25K or 5K media that contained LMK235 at the indicated doses for 12 h were collected to detect the levels of MKK7, p-JNK, p-c-Jun, c-Jun by WB, or mkk7 mRNA by Q-PCR. Mean ± SD, * P < 0.05 vs. siNC, # P < 0.05 vs. 0.1 μM LMK235.

Article Snippet: The HDAC inhibitors SAHA, M344, VPA, and TSA and the HDAC4 inhibitor LMK235 were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Inhibition, Expressing, Activity Assay, Transfection, Staining, Microscopy, Knockdown

Journal: Frontiers in Cellular Neuroscience

Article Title: Inhibition of HDAC4 Attenuated JNK/c-Jun-Dependent Neuronal Apoptosis and Early Brain Injury Following Subarachnoid Hemorrhage by Transcriptionally Suppressing MKK7

doi: 10.3389/fncel.2019.00468

Figure Lengend Snippet: Inhibition of HDAC4 rescues 5K-induced apoptosis. (A,B) CGNs were transfected with siNC, siHDAC4-a, or siHDAC4-b together with pCMV-EGFP. After 48 h, neurons were exposed to 5K media for 12 h and subjected to nucleic staining and apoptotic analysis as in . Mean ± SD, * P < 0.05 vs. 25K, # P < 0.05 vs. siHDAC4-a or siHDAC4-b. (C–F) CGNs in 25K or 5K that contained LMK235 at the indicated doses for 12 h were subjected to apoptotic analysis by nuclear staining with Hoechst 33258 or PI. Mean ± SD, * P < 0.05 vs. 25K, # P < 0.05 vs. 5K, $ P < 0.05 vs. 0.1 μM LMK235, ∧ P < 0.05 vs. 0.5 μM LMK235.

Article Snippet: The HDAC inhibitors SAHA, M344, VPA, and TSA and the HDAC4 inhibitor LMK235 were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Inhibition, Transfection, Staining

Journal: Frontiers in Cellular Neuroscience

Article Title: Inhibition of HDAC4 Attenuated JNK/c-Jun-Dependent Neuronal Apoptosis and Early Brain Injury Following Subarachnoid Hemorrhage by Transcriptionally Suppressing MKK7

doi: 10.3389/fncel.2019.00468

Figure Lengend Snippet: Administration of LMK235 significantly ameliorated the early brain injury (EBI) process with a reduction in MKK7 transcription, JNK/c-Jun activity, and neuronal apoptosis. (A,B) The levels of Ac-H3K9, H3, MKK7, p-JNK, JNK, p-c-Jun, GAPDH, or mkk7 mRNA were determined by WB or Q-PCR at 24 h post SAH in SAH + vehicle rats or SAH + LMK235 rats. Mean ± SD, * P < 0.05, n = 6 per group. (C) The p-c-Jun (ser 73) was detected by IF at 24 h post SAH, and the p-c-Jun positive rates of all NeuN-stained cells were calculated in SAH + vehicle rats or SAH + LMK235 rats; scale bar = 50 μm. Mean ± SD, * P < 0.05, n = 6 per group. (D–F) At 24 h post SAH, the differences in the apoptotic rate, neurobehavioral performance, and brain water content in the left or right hemisphere were compared between SAH + vehicle and SAH + LMK235 groups. Mean ± SD, * P < 0.05, n = 6 per group. (G) A schematic diagram illustrating the potential mechanisms involved in neuronal apoptosis regulated by HDAC4/MKK7/JNK/c-Jun axis. HDAC4 contributes to MKK7 transcription and expression, which subsequently evokes JNK/c-Jun-dependent neuronal apoptosis and EBI following SAH.

Article Snippet: The HDAC inhibitors SAHA, M344, VPA, and TSA and the HDAC4 inhibitor LMK235 were purchased from Selleck Chemicals (Shanghai, China).

Techniques: Activity Assay, Staining, Expressing