hdac5 rabbit monoclonal antibody  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: HDAC5 -encoded Microprotein NISM Mediates Nucleolar Formation and Ribosomal RNA Synthesis

doi: 10.64898/2026.02.21.707204

Figure Lengend Snippet: (A-B) BedGraph tracks of HEK293T Ribo-seq coverage across the entire HDAC5 transcript (A) and zoomed in on the NISM smORF within the 5′-UTR (B). In (A), the NISM smORF is highlighted in yellow. HDAC5 is on the negative strand and thus the 5′ to 3′ orientation runs right to left. The top tracks show read coverage for cells pre-treated with harringtonine (Harr) to capture translation initiation sites and the bottom tracks show read coverage for untreated cells lysed in the presence of cycloheximide to capture elongating ribosomes. PhyloCSF scores for each reading frame are shown below the RefSeq transcript tracks. Possible start codons in the NISM smORF are denoted by ‘M’ in (B). (C) Sequence alignment showing amino acid level conservation of NISM across distant mammals. (D) Immunoblot analysis of short and long NISM-ALFA expression in HEK293T cells. (E) ESMFold predicted structure of short NISM. Low confidence predictions are colored yellow and very low confidence predictions are colored orange. (F) Representative immunofluorescence images of NPM1 (red), FBL (cyan), and NISM-ALFA (green) in HEK293T cells transfected with empty vector (EV), short NISM-ALFA, or long NISM-ALFA for 48 h. Nuclei were counter-stained with DAPI (blue). Scale bar, 5 µm. All data are representative of at least two biological replicates.

Article Snippet: Expression of HDAC5 at the protein level was also validated by SDS-PAGE and immunoblot using the HDAC5 rabbit monoclonal antibody (Cell Signaling, 20458) following IP of HDAC5 from 500 μg of total protein from WT or NISM KO U2OS cells.

Techniques: Sequencing, Western Blot, Expressing, Immunofluorescence, Transfection, Plasmid Preparation, Staining

Journal: Immunity

Article Title: A PGE 2 -MEF2A axis enables context-dependent control of inflammatory gene expression

doi: 10.1016/j.immuni.2021.05.016

Figure Lengend Snippet: MEF2A controls IFN I induction by nucleic acids or pathogens and is targeted by PGE 2 partly via ERK5 (A) qRT-PCR analysis of Ifnb1 , Cxcl10 , or Irf1 in WT or Mef2a −/− iMacs stimulated for 4 h with poly(I:C) (red, top) or DMXAA (blue, bottom). Genotypes and IDs of the individual clones are shown. Bar plots represent mean ± SD. Data are from three biological replicates. ∗∗∗∗ p < 0.0001 (two-way ANOVA). (B) IFN-β release by WT or Mef2a −/− iMacs stimulated with poly(I:C) (red, top) or DMXAA (blue, bottom). Genotypes and IDs of the individual clones are shown. Bar plots represent mean ± SD. Data are from three biological replicates. ∗∗∗∗ p < 0.0001 (two-way ANOVA). (C) Experimental layout of in vitro infection of iMacs with VSV, BCG, or M.tb . (D) IFN-β release by WT (black) or Mef2a −/− (green) iMacs infected with VSV, BCG, or M.tb . MOI values are reported for each plot. Dot plots represent mean ± SD. Data are from two or three biological replicates. ∗∗∗∗ p < 0.0001, ∗∗ p < 0.005 (two-way ANOVA test). (E–G) qRT-PCR analysis of Ifnb1 , Cxcl10 , or Il12b in WT (black) or Mef2a −/− (green) iMacs infected with VSV (E), BCG (F), or M.tb (G). MOI values are reported for each plot. Dot plots represent mean ± SD. Data are from two or three biological replicates. ∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001 (two-way ANOVA). (H and I) Western blot analyses of HDAC5 and loading control (VCL) in BMDMs stimulated with PGE 2 for the indicated times (H) or in BMDMs upon CRISPR-Cas9-mediated targeting of Hdac5 (I). (J) qRT-PCR analysis of PGE 2 -sensitive and -resistant genes in WT (black) or HDAC5-deficient (orange) BMDMs stimulated as indicated. Dot plots represent mean ± SD. Data from three biological replicates. ∗∗∗ p < 0.001, ∗∗ p < 0.01, ∗ p < 0.05 (unpaired t test). (K) Western blot analyses for phosphorylated ERK5 (Thr218/Tyr220) as well as ERK5 and VCL as loading controls in BMDMs stimulated as indicated. (L) qRT-PCR analysis of PGE 2 -sensitive and resistant genes in WT (black) or Mapk7 −/− (orange) iMacs clones stimulated as indicated. Genotypes and IDs of the individual clones are shown. Dot plots represent mean ± SD. Data are from three biological replicates. ∗∗∗∗ p < 0.0001, ∗∗∗ p < 0.001 (unpaired t test). (M) Western blot analyses of ERK5 and VCL in WT and Mapk7 −/− iMacs clones. Genotypes and IDs of the individual clones are shown. (N) IFN-β release by WT (black) and Mapk7 −/− (orange) iMacs stimulated as indicated. Dot plot represents mean ± SD. Data are from three biological replicates. ∗∗∗∗ p < 0.0001 (two-way ANOVA). See also Figure S7 .

Article Snippet: Rabbit anti-HDAC5 (D1J7V) Monoclonal antibody , Cell Signaling Technology , Cat. 20458; RRID_ AB_2713973.

Techniques: Quantitative RT-PCR, Clone Assay, In Vitro, Infection, Western Blot, Control, CRISPR

Journal: Immunity

Article Title: A PGE 2 -MEF2A axis enables context-dependent control of inflammatory gene expression

doi: 10.1016/j.immuni.2021.05.016

Figure Lengend Snippet:

Article Snippet: Rabbit anti-HDAC5 (D1J7V) Monoclonal antibody , Cell Signaling Technology , Cat. 20458; RRID_ AB_2713973.

Techniques: Control, Virus, Recombinant, CRISPR, Produced, Plasmid Preparation, DNA Purification, Staining, Saline, Protease Inhibitor, Modification, Cell Culture, SYBR Green Assay, Reverse Transcription, TA Cloning, Sequencing, Enzyme-linked Immunosorbent Assay, Luciferase, Multiplex Assay, DNA Library Preparation, Knock-In, Software