sirt2 sirna  (Addgene inc)

Journal: Advanced Science

Article Title: Fn14 Controls the SIRT2‐Mediated Deacetylation of Slug to Inhibit the Metastasis of Epithelial Ovarian Cancer

doi: 10.1002/advs.202501552

Figure Lengend Snippet: Fn14 promotes the acetylation‐dependent degradation of Slug protein. A) Upon CHX treatment, western blot analysis detected the expression of Slug in HO8910‐PM cells after infection with Fn14 lentivirus (n = 3). B) Upon CHX treatment, western blot analysis detected the expression of Slug in ES2 cells after transfected with the Fn14 siRNA (n = 3). C) Co‐IP analysis detecting the level of ubiquitination and acetylation of Slug in HO8910‐PM and ES2 cells with specific treatment (n = 3). D) Western blot analysis was performed to detect the expression of Slug in EOC cells with selective inhibitors of Sirt family proteins (Selisistat, SIRT1inhibitor; AGK2, SIRT2 inhibitor; 3‐TYP, SIRT3 inhibitor; SIRT4‐IN‐1, SIRT4 inhibitor; SIRT5 inhibitor 4, SIRT5 inhibitor; OSS_128167, SIRT6 inhibitor; SIRT7 inhibitor 97491, SIRT7 inhibitor). E,F) Western blot analysis was performed to examine the expression of Slug in EOC cells transfected with the SIRT2 siRNA (n = 3). G) Western blot analysis was performed to examine the expression of Slug in EOC cells transfected with SIRT2 ectopically expressing plasmid(n = 3). H)Western blot analysis detecting the level of SIRT2 in HO8910‐PM and ES2 cells with up‐ or down‐regulated expression of Fn14 (n = 3). Unpaired Student's t ‐test was used to compare two groups and one‐way ANOVA was used to compare multiple groups. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SIRT2 siRNA was purchased from Santa Cruz (sc‐40988).

Techniques: Western Blot, Expressing, Infection, Transfection, Co-Immunoprecipitation Assay, Ubiquitin Proteomics, Plasmid Preparation

Journal: Advanced Science

Article Title: Fn14 Controls the SIRT2‐Mediated Deacetylation of Slug to Inhibit the Metastasis of Epithelial Ovarian Cancer

doi: 10.1002/advs.202501552

Figure Lengend Snippet: Fn14 enhances acetylation and degradation of Slug by interaction with SIRT2. A,B) Detecting the distribution of SIRT2 and Slug in cytoplasm or nucleus in EOC cells with up‐ or down‐regulated expression of Fn14 by western blot analysis (n = 3). C) Immunofluorescence of Fn14 (red) and SIRT2 (green) in HO8910‐PM cells infected with Fn14 lentivirus. DAPI (blue) was used to visualize the nucleus (Scale bar = 20 µm). D) Immunofluorescence of Slug (red) and SIRT2 (green) in HO8910‐PM cells infected with Fn14 lentivirus (Scale bar = 20 µm). E) Lysates from HEK293T cells transfected with Fn14 and SIRT2 were subjected to anti‐Fn14 or anti‐SIRT2 immunoprecipitation. Immunoblots of Fn14 and SIRT2 are shown (n = 3). F) Lysates from HO8910‐PM and ES2 with w‐specific treatment were subjected to anti‐SIRT2 immunoprecipitation. Immunoblots of endogenous Fn14 and Slug are shown (n = 3). G) Interaction between recombinant Fn14 and SIRT2 was detected by pull‐down analysis (n = 3). H) Lysates from HEK293T cells transfected with GFP‐tagged truncation of Fn14 and SIRT2 were subjected to anti‐SIRT2 immunoprecipitation. Immunoblots of GFP are shown (n = 3). Unpaired Student's t‐test was used to compare two groups, one‐way ANOVA was used to compare multiple groups. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SIRT2 siRNA was purchased from Santa Cruz (sc‐40988).

Techniques: Expressing, Western Blot, Immunofluorescence, Infection, Transfection, Immunoprecipitation, Recombinant

Journal: Advanced Science

Article Title: Fn14 Controls the SIRT2‐Mediated Deacetylation of Slug to Inhibit the Metastasis of Epithelial Ovarian Cancer

doi: 10.1002/advs.202501552

Figure Lengend Snippet: Silencing SIRT2 inhibits the invasion and metastasis of EOC cells. A–D) Cell migration and invasion were examined by wound healing assay and transwell assay in HO8910‐PM and ES2 cells transfected with the SIRT2 siRNA (n = 3 per cell line, Transwell scale bar = 100 µm, Wound healing scale bar = 200 µm). E–H) Western blot analysis detecting the EMT makers in HO8910‐PM and ES2 cells with transfected with the SIRT2 siRNA(n = 3). Unpaired Student's t ‐test was used to compare two groups, one‐way ANOVA was used to compare multiple groups, and Chi‐square test was applied for comparisons of proportions. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SIRT2 siRNA was purchased from Santa Cruz (sc‐40988).

Techniques: Migration, Wound Healing Assay, Transwell Assay, Transfection, Western Blot

Journal: Advanced Science

Article Title: Fn14 Controls the SIRT2‐Mediated Deacetylation of Slug to Inhibit the Metastasis of Epithelial Ovarian Cancer

doi: 10.1002/advs.202501552

Figure Lengend Snippet: Fn14 alleviates the metastasis of EOC cells in vivo. A) Body weight of mice with HO8910(HO+DMSO), HO8910‐PM/Ctrl (PM‐Ctrl+DMSO), HO8910‐PM/Fn14(PM‐Fn14+DMSO), HO8910‐PM/shSlug (PM‐shslug+DMSO), HO8910‐PM/AGK2(PM‐Ctrl+AGK2) implant are shown. B,C) Metastasis of tumor in five groups are shown. D) Representative hematoxylin–eosin staining and summarized data on lung foci in five groups. E) Tumors of each group were immunohistochemically tested for Fn14, SIRT2, and Slug. One‐way ANOVA was used to compare multiple groups and the Chi‐square test was applied for comparisons of proportions. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SIRT2 siRNA was purchased from Santa Cruz (sc‐40988).

Techniques: In Vivo, Staining

Journal: Advanced Science

Article Title: Fn14 Controls the SIRT2‐Mediated Deacetylation of Slug to Inhibit the Metastasis of Epithelial Ovarian Cancer

doi: 10.1002/advs.202501552

Figure Lengend Snippet: Fn14‐SIRT2‐Slug axis correlates with the prognosis of EOC patients. A) IHC was performed to detect the expression of Fn14, SIRT2, and Slug in EOC samples. B) Spearman's correlation analysis was performed to examine the correlation between the expression levels of Fn14, SIRT2, and Slug in EOC samples. C,D)The correlation between SIRT2 and Slug expression and patient survival was demonstrated in the survival analysis. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SIRT2 siRNA was purchased from Santa Cruz (sc‐40988).

Techniques: Expressing

Journal: Advanced Science

Article Title: Fn14 Controls the SIRT2‐Mediated Deacetylation of Slug to Inhibit the Metastasis of Epithelial Ovarian Cancer

doi: 10.1002/advs.202501552

Figure Lengend Snippet: Graphical abstract of Fn14 controls the SIRT2‐mediated deacetylation of Slug to inhibit the metastasis of EOC. Fn14 inhibited EOC metastasis by regulating Slug‐mediated EMT. Furthermore, Fn14 altered the subcellular localization of SIRT2 by interacting with SIRT2, leading to reduced SIRT2 shuttling into the nucleus and subsequently promoting the acetylated degradation of Slug.

Article Snippet: SIRT2 siRNA was purchased from Santa Cruz (sc‐40988).

Techniques:

Journal: Journal of Inflammation Research

Article Title: SIRT2 Promotes NLRP3-Mediated Microglia Pyroptosis and Neuroinflammation via FOXO3a Pathway After Subarachnoid Hemorrhage

doi: 10.2147/JIR.S487716

Figure Lengend Snippet: SIRT2 activates NLRP3 inflammasome-induced microglial pyroptosis by activating FOXO3a and promoting its nuclear localization. ( A ) Western blot analysis and quantification of FOXO3a protein expression in Sham, SAH, Si-SIRT2+Sham, and Si-SIRT2+SAH groups. ( B ) Western blot analysis and quantification of FOXO3a protein expression in Control, Hb, Si-SIRT2+Control, and Si-SIRT2+Hb groups. ( C ) Nucleoplasmic separation assay to observe the effect of siRNA knockdown of SIRT2 post-SAH on FOXO3a protein expression levels in the cytoplasm and nucleus of microglia. ( D ) Immunofluorescence analysis of FOXO3a distribution in the nucleus and cytoplasm of microglial cells. Data are in mean ± SD, * P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: According to the manufacturer’s instructions and siRNA efficacy assessment, SIRT2 siRNA (siB14529151009-1-5, Ribobio) was administered at a rate of 0.5 µL/min into the left lateral ventricle two days prior to SAH induction.

Techniques: Western Blot, Expressing, Control, Knockdown, Immunofluorescence