hdac6 enzymatic activity Search Results


human hdac6  (Cell Signaling Technology Inc)
95
Cell Signaling Technology Inc human hdac6
(A) Workflow for evaluating NLRP3 inflammasome activation in BMDMs. Bone marrow was isolated from mice strains with different mutations in <t>HDAC6</t> as indicated and BMDMs were expanded for 7 days in the presence of M-CSF1. Subsequently, BMDMs were primed for 4 hrs with LPS and NLRP3 inflammasome formation was induced by nigericin or MSU addition. Release of IL-1β was monitored by ELISA and activation/cleavage of Caspase-1 was evaluated by immunoblotting. (B) Normal NLRP3 activation in WT, HDAC6 KO and HDAC6 ZnF m BMDMs. Immunoblot monitoring HDAC6, Capase-1 (p20) cleavage and α-tubulin as loading control. nigericin and MSU were used with LPS primed BMDMs (n = 3). (C) Normal mIL-1β release in the supernatant from WT, HDAC6 KO and ZnF m BMDMs. IL-1β from (B) was measured by ELISA and statistical analysis was done with one-way ANOVA. ns, no significant difference. (D) No difference in cytotoxicity following inflammasome induction in WT, HDAC6 KO and ZnF m BMDMs. Lactate dehydrogenase (LDH) was measured in the culture supernatant from (B) and statistical analysis was done by one-way ANOVA. ns, no significant difference.
Human Hdac6, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/hdac6+enzymatic+activity/HDAC6+Rabbit+mAb/bio_rxiv__2023__08__15__553363-115-8-10
Average 95 stars, based on 1 article reviews
human hdac6 - by Bioz Stars, 2026-10
95/100 stars
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live cell analysis instruments  (Sartorius AG)
99
Sartorius AG live cell analysis instruments
(A) Workflow for evaluating NLRP3 inflammasome activation in BMDMs. Bone marrow was isolated from mice strains with different mutations in <t>HDAC6</t> as indicated and BMDMs were expanded for 7 days in the presence of M-CSF1. Subsequently, BMDMs were primed for 4 hrs with LPS and NLRP3 inflammasome formation was induced by nigericin or MSU addition. Release of IL-1β was monitored by ELISA and activation/cleavage of Caspase-1 was evaluated by immunoblotting. (B) Normal NLRP3 activation in WT, HDAC6 KO and HDAC6 ZnF m BMDMs. Immunoblot monitoring HDAC6, Capase-1 (p20) cleavage and α-tubulin as loading control. nigericin and MSU were used with LPS primed BMDMs (n = 3). (C) Normal mIL-1β release in the supernatant from WT, HDAC6 KO and ZnF m BMDMs. IL-1β from (B) was measured by ELISA and statistical analysis was done with one-way ANOVA. ns, no significant difference. (D) No difference in cytotoxicity following inflammasome induction in WT, HDAC6 KO and ZnF m BMDMs. Lactate dehydrogenase (LDH) was measured in the culture supernatant from (B) and statistical analysis was done by one-way ANOVA. ns, no significant difference.
Live Cell Analysis Instruments, supplied by Sartorius AG, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/hdac6+enzymatic+activity/Live+Cell+Analysis+Instruments/sartorius+ag___live-cell-analysis-instruments
Average 99 stars, based on 1 article reviews
live cell analysis instruments - by Bioz Stars, 2026-10
99/100 stars
  Buy from Supplier

Image Search Results


(A) Workflow for evaluating NLRP3 inflammasome activation in BMDMs. Bone marrow was isolated from mice strains with different mutations in HDAC6 as indicated and BMDMs were expanded for 7 days in the presence of M-CSF1. Subsequently, BMDMs were primed for 4 hrs with LPS and NLRP3 inflammasome formation was induced by nigericin or MSU addition. Release of IL-1β was monitored by ELISA and activation/cleavage of Caspase-1 was evaluated by immunoblotting. (B) Normal NLRP3 activation in WT, HDAC6 KO and HDAC6 ZnF m BMDMs. Immunoblot monitoring HDAC6, Capase-1 (p20) cleavage and α-tubulin as loading control. nigericin and MSU were used with LPS primed BMDMs (n = 3). (C) Normal mIL-1β release in the supernatant from WT, HDAC6 KO and ZnF m BMDMs. IL-1β from (B) was measured by ELISA and statistical analysis was done with one-way ANOVA. ns, no significant difference. (D) No difference in cytotoxicity following inflammasome induction in WT, HDAC6 KO and ZnF m BMDMs. Lactate dehydrogenase (LDH) was measured in the culture supernatant from (B) and statistical analysis was done by one-way ANOVA. ns, no significant difference.

Journal: bioRxiv

Article Title: HDAC6/aggresome processing pathway importance for inflammasome formation is context dependent

doi: 10.1101/2023.08.15.553363

Figure Lengend Snippet: (A) Workflow for evaluating NLRP3 inflammasome activation in BMDMs. Bone marrow was isolated from mice strains with different mutations in HDAC6 as indicated and BMDMs were expanded for 7 days in the presence of M-CSF1. Subsequently, BMDMs were primed for 4 hrs with LPS and NLRP3 inflammasome formation was induced by nigericin or MSU addition. Release of IL-1β was monitored by ELISA and activation/cleavage of Caspase-1 was evaluated by immunoblotting. (B) Normal NLRP3 activation in WT, HDAC6 KO and HDAC6 ZnF m BMDMs. Immunoblot monitoring HDAC6, Capase-1 (p20) cleavage and α-tubulin as loading control. nigericin and MSU were used with LPS primed BMDMs (n = 3). (C) Normal mIL-1β release in the supernatant from WT, HDAC6 KO and ZnF m BMDMs. IL-1β from (B) was measured by ELISA and statistical analysis was done with one-way ANOVA. ns, no significant difference. (D) No difference in cytotoxicity following inflammasome induction in WT, HDAC6 KO and ZnF m BMDMs. Lactate dehydrogenase (LDH) was measured in the culture supernatant from (B) and statistical analysis was done by one-way ANOVA. ns, no significant difference.

Article Snippet: Antibody information: mouse Caspase-1 (p20) (AdipoGen#AG-20B-0042-C100), mHDAC6 (homemade), human HDAC6 (CellSignaling#7558), α-tubulin (Sigma-Aldrich #T9026), HA (CellSignaling#3724), pan Actin (Sigma-Aldrich#SAB4502632).

Techniques: Activation Assay, Isolation, Enzyme-linked Immunosorbent Assay, Western Blot, Control

(A) Structure of the DARPin F10 - HDAC6 ZnF domain complex (PDB: 7ZYU) showing blockade of Ub recruitment. The surface representation shows HDAC6 ZnF (yellow) and the C-terminal LRGG of Ub (light pink) (overlayed with PDB:3GV4), the ribbon representation depicts DARPIN F10 (cyan). (B) Establishment of a THP-1 F10-FKBP cell line with degradable DARPin F10. Immunoblotting with lysates of THP-1 F10-FKBP cells treated at the indicated concentration with dTAG (for 6 hrs). The leftmost lane (WT) shows the parental THP-1 cells. The membrane was probed with antibodies against actin as loading control and F10 (HA-F10-FKBP F36V , detected with anti-HA). (C) IL-1β release under various treatments in THP-1 F10-FKBP cells at 3 hrs post-nigericin stimulation. Cells were treated as indicated in the scheme at the top. The graph at the bottom shows the measurement of IL-1β in cell supernatants, based on 3 independent experiments. MCC950 and CGP084892 are NLRP3 and Caspase-1 inhibitors, respectively. One-way ANOVA test was applied for statistical analysis. ****, p<0.0001. (D) IL-1β release from 0 hr to 3.5 hr after nigericin treatment in THP-1 or THP-1-F10-FKBP cells. Experimental set-up was as in (B) and IL-1β concentration was monitored at timepoint 0, 0.5, 1.5, and 3.5 hr post nigericin treatment. Two-way ANOVA tests were applied, and comparison was performed within each timepoint between with and without dTAG group. **, p<0.01.

Journal: bioRxiv

Article Title: HDAC6/aggresome processing pathway importance for inflammasome formation is context dependent

doi: 10.1101/2023.08.15.553363

Figure Lengend Snippet: (A) Structure of the DARPin F10 - HDAC6 ZnF domain complex (PDB: 7ZYU) showing blockade of Ub recruitment. The surface representation shows HDAC6 ZnF (yellow) and the C-terminal LRGG of Ub (light pink) (overlayed with PDB:3GV4), the ribbon representation depicts DARPIN F10 (cyan). (B) Establishment of a THP-1 F10-FKBP cell line with degradable DARPin F10. Immunoblotting with lysates of THP-1 F10-FKBP cells treated at the indicated concentration with dTAG (for 6 hrs). The leftmost lane (WT) shows the parental THP-1 cells. The membrane was probed with antibodies against actin as loading control and F10 (HA-F10-FKBP F36V , detected with anti-HA). (C) IL-1β release under various treatments in THP-1 F10-FKBP cells at 3 hrs post-nigericin stimulation. Cells were treated as indicated in the scheme at the top. The graph at the bottom shows the measurement of IL-1β in cell supernatants, based on 3 independent experiments. MCC950 and CGP084892 are NLRP3 and Caspase-1 inhibitors, respectively. One-way ANOVA test was applied for statistical analysis. ****, p<0.0001. (D) IL-1β release from 0 hr to 3.5 hr after nigericin treatment in THP-1 or THP-1-F10-FKBP cells. Experimental set-up was as in (B) and IL-1β concentration was monitored at timepoint 0, 0.5, 1.5, and 3.5 hr post nigericin treatment. Two-way ANOVA tests were applied, and comparison was performed within each timepoint between with and without dTAG group. **, p<0.01.

Article Snippet: Antibody information: mouse Caspase-1 (p20) (AdipoGen#AG-20B-0042-C100), mHDAC6 (homemade), human HDAC6 (CellSignaling#7558), α-tubulin (Sigma-Aldrich #T9026), HA (CellSignaling#3724), pan Actin (Sigma-Aldrich#SAB4502632).

Techniques: Western Blot, Concentration Assay, Membrane, Control, Comparison

(A) Schematic of the mode of action of an HDAC6 degrader. The PROTAC engages both HDAC6 and the E3 ligase cereblon (CRBN), which then leads to ubiquitination and degradation of HDAC6. (B) Chemical structure of the HDAC6 degrader XY-07-35 and of the negative control XY-07-191 which no longer recruits CRBN. (C) In vitro enzymatic assay of compound XY-07-35 on the activity of different HDACs. IC 50 for each enzyme is shown. (D) XY-07-35 mediated HDAC6 degradation in PBMCs. PBMCs (without or with LPS priming for 4 hrs) were treated with the indicated compounds and HDAC6 levels were detected by immunoblotting with an anti-HDAC6 antibody. Detection of actin was used as control for loading. CGP, CGP084892 (Caspase-1 inhibitor); AFN700 is an inhibitor of the NF-kB pathway (IKK inhibitor). (E) HDAC6 degradation impact on IL-1β release in PBMCs following MSU, nigericin or ATP treatment. The scheme at the top depicts the experimental outline. The graphs below show the results of 3 independent experiments using the inducers as indicated. NP3-948 is an inhibitor of the NLRP3 inflammasome.

Journal: bioRxiv

Article Title: HDAC6/aggresome processing pathway importance for inflammasome formation is context dependent

doi: 10.1101/2023.08.15.553363

Figure Lengend Snippet: (A) Schematic of the mode of action of an HDAC6 degrader. The PROTAC engages both HDAC6 and the E3 ligase cereblon (CRBN), which then leads to ubiquitination and degradation of HDAC6. (B) Chemical structure of the HDAC6 degrader XY-07-35 and of the negative control XY-07-191 which no longer recruits CRBN. (C) In vitro enzymatic assay of compound XY-07-35 on the activity of different HDACs. IC 50 for each enzyme is shown. (D) XY-07-35 mediated HDAC6 degradation in PBMCs. PBMCs (without or with LPS priming for 4 hrs) were treated with the indicated compounds and HDAC6 levels were detected by immunoblotting with an anti-HDAC6 antibody. Detection of actin was used as control for loading. CGP, CGP084892 (Caspase-1 inhibitor); AFN700 is an inhibitor of the NF-kB pathway (IKK inhibitor). (E) HDAC6 degradation impact on IL-1β release in PBMCs following MSU, nigericin or ATP treatment. The scheme at the top depicts the experimental outline. The graphs below show the results of 3 independent experiments using the inducers as indicated. NP3-948 is an inhibitor of the NLRP3 inflammasome.

Article Snippet: Antibody information: mouse Caspase-1 (p20) (AdipoGen#AG-20B-0042-C100), mHDAC6 (homemade), human HDAC6 (CellSignaling#7558), α-tubulin (Sigma-Aldrich #T9026), HA (CellSignaling#3724), pan Actin (Sigma-Aldrich#SAB4502632).

Techniques: Ubiquitin Proteomics, Negative Control, In Vitro, Enzymatic Assay, Activity Assay, Western Blot, Control

THP-1 cells, with or without in vitro differentiation, were incubated with XY-07-35 and the HDAC6 level was examined by immunoblotting.

Journal: bioRxiv

Article Title: HDAC6/aggresome processing pathway importance for inflammasome formation is context dependent

doi: 10.1101/2023.08.15.553363

Figure Lengend Snippet: THP-1 cells, with or without in vitro differentiation, were incubated with XY-07-35 and the HDAC6 level was examined by immunoblotting.

Article Snippet: Antibody information: mouse Caspase-1 (p20) (AdipoGen#AG-20B-0042-C100), mHDAC6 (homemade), human HDAC6 (CellSignaling#7558), α-tubulin (Sigma-Aldrich #T9026), HA (CellSignaling#3724), pan Actin (Sigma-Aldrich#SAB4502632).

Techniques: In Vitro, Incubation, Western Blot

(A) XY-07-35 mediated HDAC6 degradation in pyrin-expressing U937 cells. Cells were treated with the indicated compounds and HDAC6 was detected by immunoblotting with an anti-HDAC6 antibody. Detection of actin was used as a control for protein loading. (B) No impact of HDAC6 degradation on IL-18 release in U937-pyrin cells treated with the indicated compounds. The scheme at the top depicts the experimental outline. The graph at the bottom presents IL-1β release based on 4 independent experiments. CGP, CGP084892.

Journal: bioRxiv

Article Title: HDAC6/aggresome processing pathway importance for inflammasome formation is context dependent

doi: 10.1101/2023.08.15.553363

Figure Lengend Snippet: (A) XY-07-35 mediated HDAC6 degradation in pyrin-expressing U937 cells. Cells were treated with the indicated compounds and HDAC6 was detected by immunoblotting with an anti-HDAC6 antibody. Detection of actin was used as a control for protein loading. (B) No impact of HDAC6 degradation on IL-18 release in U937-pyrin cells treated with the indicated compounds. The scheme at the top depicts the experimental outline. The graph at the bottom presents IL-1β release based on 4 independent experiments. CGP, CGP084892.

Article Snippet: Antibody information: mouse Caspase-1 (p20) (AdipoGen#AG-20B-0042-C100), mHDAC6 (homemade), human HDAC6 (CellSignaling#7558), α-tubulin (Sigma-Aldrich #T9026), HA (CellSignaling#3724), pan Actin (Sigma-Aldrich#SAB4502632).

Techniques: Expressing, Western Blot, Control