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rac1 inhibitor w56  (MedChemExpress)
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MedChemExpress rac1 inhibitor w56
SIRT7 induces cytoskeletal remodeling via <t>RAC1.</t> ( A ) RAC1activation in H37Rv-infected Sirt7 −/− and Sirt7 +/+ BMDMs assayed at 0 min and 30 min post-infection. “Input” represents RAC1 protein levels in whole-cell lysates, while “IP: PAK1-PBD” indicates active RAC1 levels. ( B ) Immunoblot and immunoprecipitation assays illustrating the interaction of SIRT7 and RAC1 in THP1-derived macrophages transfected with si- RAC1 or normal control (si-NC) siRNA. ( C and G ) Flow cytometry analysis quantifying the phagocytosis of GFP-H37Ra by Sirt7 −/− and Sirt7 +/+ BMDMs under the conditions of RAC1 inhibitor or activator treatment. GFP + BMDMs represent cells that phagocytosed GFP-H37Ra. ( D ) Intracellular CFU counts of H37Rv at 4 hours post-infection in Sirt7 −/− and Sirt7 +/+ BMDMs. ( E and H ) Following a 4-hour infection period with H37Rv, macrophages were washed with phosphate-buffered saline (PBS) to eliminate unphagocytosed bacteria. Subsequently, a fresh complete medium was introduced, either containing an RAC1 inhibitor or activator. CFU counts were determined 68 hours later. ( F and I ) The survival ratio of H37Rv was evaluated in Sirt7 −/− BMDMs compared to WT BMDMs, following treatment with either RAC1 inhibitor or activator. ( J and K ) Immunoblot analysis was performed to assess the expression of LC3II in Sirt7 −/− and Sirt7 +/+ BMDMs treated with an RAC1 inhibitor or Baf-A1. The expression levels of LC3II were quantified and are shown relative to GAPDH, as indicated below the corresponding bands. Data are presented as means ± SEM, * P < 0.05, ** P < 0.01, and *** P < 0.001, as determined by one-way ANOVA with Tukey’s multiple comparisons test. Each experiment was independently replicated two to three times.
Rac1 Inhibitor W56, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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SIRT7 induces cytoskeletal remodeling via RAC1. ( A ) RAC1activation in H37Rv-infected Sirt7 −/− and Sirt7 +/+ BMDMs assayed at 0 min and 30 min post-infection. “Input” represents RAC1 protein levels in whole-cell lysates, while “IP: PAK1-PBD” indicates active RAC1 levels. ( B ) Immunoblot and immunoprecipitation assays illustrating the interaction of SIRT7 and RAC1 in THP1-derived macrophages transfected with si- RAC1 or normal control (si-NC) siRNA. ( C and G ) Flow cytometry analysis quantifying the phagocytosis of GFP-H37Ra by Sirt7 −/− and Sirt7 +/+ BMDMs under the conditions of RAC1 inhibitor or activator treatment. GFP + BMDMs represent cells that phagocytosed GFP-H37Ra. ( D ) Intracellular CFU counts of H37Rv at 4 hours post-infection in Sirt7 −/− and Sirt7 +/+ BMDMs. ( E and H ) Following a 4-hour infection period with H37Rv, macrophages were washed with phosphate-buffered saline (PBS) to eliminate unphagocytosed bacteria. Subsequently, a fresh complete medium was introduced, either containing an RAC1 inhibitor or activator. CFU counts were determined 68 hours later. ( F and I ) The survival ratio of H37Rv was evaluated in Sirt7 −/− BMDMs compared to WT BMDMs, following treatment with either RAC1 inhibitor or activator. ( J and K ) Immunoblot analysis was performed to assess the expression of LC3II in Sirt7 −/− and Sirt7 +/+ BMDMs treated with an RAC1 inhibitor or Baf-A1. The expression levels of LC3II were quantified and are shown relative to GAPDH, as indicated below the corresponding bands. Data are presented as means ± SEM, * P < 0.05, ** P < 0.01, and *** P < 0.001, as determined by one-way ANOVA with Tukey’s multiple comparisons test. Each experiment was independently replicated two to three times.

Journal: mBio

Article Title: SIRT7 remodels the cytoskeleton via RAC1 to enhance host resistance to Mycobacterium tuberculosis

doi: 10.1128/mbio.00756-24

Figure Lengend Snippet: SIRT7 induces cytoskeletal remodeling via RAC1. ( A ) RAC1activation in H37Rv-infected Sirt7 −/− and Sirt7 +/+ BMDMs assayed at 0 min and 30 min post-infection. “Input” represents RAC1 protein levels in whole-cell lysates, while “IP: PAK1-PBD” indicates active RAC1 levels. ( B ) Immunoblot and immunoprecipitation assays illustrating the interaction of SIRT7 and RAC1 in THP1-derived macrophages transfected with si- RAC1 or normal control (si-NC) siRNA. ( C and G ) Flow cytometry analysis quantifying the phagocytosis of GFP-H37Ra by Sirt7 −/− and Sirt7 +/+ BMDMs under the conditions of RAC1 inhibitor or activator treatment. GFP + BMDMs represent cells that phagocytosed GFP-H37Ra. ( D ) Intracellular CFU counts of H37Rv at 4 hours post-infection in Sirt7 −/− and Sirt7 +/+ BMDMs. ( E and H ) Following a 4-hour infection period with H37Rv, macrophages were washed with phosphate-buffered saline (PBS) to eliminate unphagocytosed bacteria. Subsequently, a fresh complete medium was introduced, either containing an RAC1 inhibitor or activator. CFU counts were determined 68 hours later. ( F and I ) The survival ratio of H37Rv was evaluated in Sirt7 −/− BMDMs compared to WT BMDMs, following treatment with either RAC1 inhibitor or activator. ( J and K ) Immunoblot analysis was performed to assess the expression of LC3II in Sirt7 −/− and Sirt7 +/+ BMDMs treated with an RAC1 inhibitor or Baf-A1. The expression levels of LC3II were quantified and are shown relative to GAPDH, as indicated below the corresponding bands. Data are presented as means ± SEM, * P < 0.05, ** P < 0.01, and *** P < 0.001, as determined by one-way ANOVA with Tukey’s multiple comparisons test. Each experiment was independently replicated two to three times.

Article Snippet: The following chemicals were used: RAC1 Inhibitor W56 (Medchemexpress, #HY-P1382), RAC/CDC42 Activator II (cytoskeleton, #CN02), pHrodo Red succinimidyl (NHS) ester (Invitrogen, #P36600), Mouse Intracellular (nuclear/transcription factor) Protein Flow Cytometry Workflow Kit (Invitrogen, # A53017), pHrodo Red succinimidyl ester (Invitrogen, #P36600), Active Rac1 Pull-Down and Detection Kit (Thermo Scientific, #16118), SPHEROTM Fluorescent Particles (Spherotech, #FL-6068), Bafilomycin A1 (MCE, #HY-100558), Chloroquine (MCE, # HY-17589A), fluorescein isothiocyanate (FITC) Annexin V Apoptosis Detection Kit with PI (BioLegend, #640914), MitoSOX Mitochondrial Superoxide Indicators (Invitrogen, # M36008), H2DCF-DA (Invitrogen, # D399), Mouse CXCL1 ELISA Kit (HUABIO, # EM0017), Mouse CXCL2 ELISA Kit (HUABIO, # EM0018).

Techniques: Infection, Western Blot, Immunoprecipitation, Derivative Assay, Transfection, Control, Flow Cytometry, Saline, Bacteria, Expressing

Overexpression of Sirt7 enhanced host anti-TB immunity. ( A and B ) Intracellular CFU counts of H37Rv at 4 hours and 72 hours post-infection in Sirt7 +/+ and Sirt7 TG BMDMs. ( C ) The survival ratio of H37Rv was evaluated in Sirt7 TG BMDMs compared to WT controls. ( D ) LC3II expression was elevated in Sirt7 TG BMDMs compared to WT controls. ( E ) RAC1 activation in H37Rv-infected Sirt7 +/+ and Sirt7 TG BMDMs assayed at 0 min and 30 min post-infection, with “Input” indicating total RAC1 and “IP: PAK1-PBD” indicates active RAC1. ( F and G ) Bacterial burden in the lungs and spleens of Sirt7 +/+ and Sirt7 TG mice was assessed in the tissue homogenates. ( H ) Histopathological analysis of lung sections from Sirt7 +/+ and Sirt7 TG mice infected with H37Rv. Sections were stained with hematoxylin and eosin, and images were captured using a NanoZoomer digital pathology system. Data are presented as means ± SEM, * P < 0.05, ** P < 0.01, and *** P < 0.001 by Student’s two-tailed unpaired t-test. Each experiment was independently replicated two to three times.

Journal: mBio

Article Title: SIRT7 remodels the cytoskeleton via RAC1 to enhance host resistance to Mycobacterium tuberculosis

doi: 10.1128/mbio.00756-24

Figure Lengend Snippet: Overexpression of Sirt7 enhanced host anti-TB immunity. ( A and B ) Intracellular CFU counts of H37Rv at 4 hours and 72 hours post-infection in Sirt7 +/+ and Sirt7 TG BMDMs. ( C ) The survival ratio of H37Rv was evaluated in Sirt7 TG BMDMs compared to WT controls. ( D ) LC3II expression was elevated in Sirt7 TG BMDMs compared to WT controls. ( E ) RAC1 activation in H37Rv-infected Sirt7 +/+ and Sirt7 TG BMDMs assayed at 0 min and 30 min post-infection, with “Input” indicating total RAC1 and “IP: PAK1-PBD” indicates active RAC1. ( F and G ) Bacterial burden in the lungs and spleens of Sirt7 +/+ and Sirt7 TG mice was assessed in the tissue homogenates. ( H ) Histopathological analysis of lung sections from Sirt7 +/+ and Sirt7 TG mice infected with H37Rv. Sections were stained with hematoxylin and eosin, and images were captured using a NanoZoomer digital pathology system. Data are presented as means ± SEM, * P < 0.05, ** P < 0.01, and *** P < 0.001 by Student’s two-tailed unpaired t-test. Each experiment was independently replicated two to three times.

Article Snippet: The following chemicals were used: RAC1 Inhibitor W56 (Medchemexpress, #HY-P1382), RAC/CDC42 Activator II (cytoskeleton, #CN02), pHrodo Red succinimidyl (NHS) ester (Invitrogen, #P36600), Mouse Intracellular (nuclear/transcription factor) Protein Flow Cytometry Workflow Kit (Invitrogen, # A53017), pHrodo Red succinimidyl ester (Invitrogen, #P36600), Active Rac1 Pull-Down and Detection Kit (Thermo Scientific, #16118), SPHEROTM Fluorescent Particles (Spherotech, #FL-6068), Bafilomycin A1 (MCE, #HY-100558), Chloroquine (MCE, # HY-17589A), fluorescein isothiocyanate (FITC) Annexin V Apoptosis Detection Kit with PI (BioLegend, #640914), MitoSOX Mitochondrial Superoxide Indicators (Invitrogen, # M36008), H2DCF-DA (Invitrogen, # D399), Mouse CXCL1 ELISA Kit (HUABIO, # EM0017), Mouse CXCL2 ELISA Kit (HUABIO, # EM0018).

Techniques: Over Expression, Infection, Expressing, Activation Assay, Staining, Two Tailed Test