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e 06 aha 4032 50s ribosomal protein l11 93 shewanella ana  (ATCC)
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ATCC e 06 aha 4032 50s ribosomal protein l11 93 shewanella ana
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sample, human serum, pediatric tanner stage i  (Golden West Biologicals)
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ex527 selleck cat  (Selleck Chemicals)
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Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM <t>EX527</t> was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.
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bio plex 200  (Bio-Rad)
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Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM <t>EX527</t> was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.
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α phosphorylated histone h3  (Cell Signaling Technology Inc)
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Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM <t>EX527</t> was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.
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polyclonal goat anti mouse igm  (SouthernBiotech)
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Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM <t>EX527</t> was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.
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nivestim  (Hospira)
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Hospira nivestim
Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM <t>EX527</t> was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.
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minutetm total protein extraction kit  (Invent Biotechnologies)
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Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM <t>EX527</t> was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.
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Image Search Results


Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM EX527 was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.

Journal: Cell reports

Article Title: LARP7 ameliorates cellular senescence and aging by allosterically enhancing SIRT1 deacetylase activity.

doi: 10.1016/j.celrep.2021.110038

Figure Lengend Snippet: Figure 2. The declined SIRT1 deacetylase activity accounted for LARP7-depletion-induced senescence (A) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 3 days. (B) Western blotting assessing the senescent regulators in IMR90 receiving LARP7 shRNA knockdown for 2 weeks. (C) LARP7 knockdown for 3 and 14 days in IMR90 cells increased the p53 K382Ac and p65 K310Ac level. The control immunoglobulin G (IgG) (rabbit for p65 and mouse for p53) didn’t pull down either p53 or p65 indicating the specificity of immunoprecipitation. (D) Knocking down LARP7 suppressed the nuclear SIRT activity on days 3 and 14. 10 mM EX527 was used as a positive control. n = 3. (E) The increased p53 and p65 acetylation upon LARP7 depletion was blocked by SRT1720. shCtrl- or shLARP7-transfected IMR90 cells were treated with 1 mM SRT1720 for 12 h and then subjected to immunoprecipitation. (F) qRT-PCR showing SRT1720 abrogated the SASP genes activation. IMR90 cells were treated with 2 mM JSH-23 for 48 h as a positive control. n = 3.

Article Snippet: N/A Bacterial and virus strains BL21/DE3 E. coli NEB Cat #C2527 Biological samples Human coronary artery samples The University of Pennsylvania Human Heart Tissue Bank IRB protocol number: 802781 Chemicals, peptides, and recombinant proteins Tamoxifen Sigma Cat #T5648; CAS:10540-29-1 SRT1720 Selleck Cat #S1129; CAS: 1001645-58-4 KU60019 Selleck Cat #S1570; CAS: 925701-49-1 Oil Red Powder Sigma Cat #O0625; CAS: 1320-06-5 EX527 Selleck Cat #S1541; CAS: 49843-98-3 Fetal bovine serum ExCell Bio Cat #FSP500 Acetyl-CoA Sigma Cat #A2181; CAS: 32140-51-5 NAD+ Selleck Cat #S2518; CAS: 53-84-9 Critical commercial assays Fast Mutagenesis System kit TransGen Biotech Cat #FM111S Senescence b-Galactosidase Staining Kit Cell Signaling Technology Cat #9860S NAD/NADH Quantification Kit Sigma Cat #MAK037 Universal SIRT Activity Assay Kit Abcam Cat #ab156915 Deposited data RNA-Seq This paper GSE160279 Processed RNA-Seq data This paper Table S1 (Continued on next page) e1 Cell Reports 37, 110038, November 23, 2021

Techniques: Histone Deacetylase Assay, Activity Assay, Western Blot, shRNA, Knockdown, Control, Immunoprecipitation, Positive Control, Transfection, Quantitative RT-PCR, Activation Assay