Journal: Aging Cell

Article Title: SIRT1 Silences L1 Retrotransposons by Stabilizing Heterochromatin‐Modifying Complexes

doi: 10.1111/acel.70689

Figure Lengend Snippet: SIRT1 suppresses cellular senescence and this effect is rescued by 3TC treatment. (A) Schematic of induction in control, SIRT1‐overexpressing and SIRT1 knockdown cells post‐IR. (B) Western blot analysis of SIRT1 expression in HCA2‐hTERT cells. The HCA2‐hTERT cells were transfected with the control vector or the SIRT1 overexpression vector. (C) Quantification of SA‐β‐gal positive HCA2‐hTERT cells. HCA2‐hTERT cells were exposed to 10 Gy of X‐ray irradiation and transfected with the control vector or the SIRT1 overexpression vector for 7 days. Images from different areas of the plate were taken and quantified. (D) Representative images of SA‐β‐gal staining in HCA2‐hTERT cells, which were transfected with the control vector or the SIRT1 overexpression vector (Scale bar: 50 μm). (E) Western blot analysis of p21 and p16 in control and SIRT1‐overexpressing HCA2‐hTERT cells. (F) Western blot analysis of SIRT1 expression in control and SIRT1‐knockdown HCA2‐hTERT cells. (G) Western blot analysis of p21 and p16 in control and SIRT1‐knockdown HCA2‐hTERT cells. (H) Relative mRNA levels of CCL2 and MMP3 in control and SIRT1‐overexpressing HCA2‐hTERT cells. (I) Relative mRNA levels of CCL2 and MMP3 in control and SIRT1‐knockdown HCA2‐hTERT cells. The cells were treated with DMSO or 3TC. H, I: bars represent mean ± SD of biological replicates. Two‐tailed Student's t ‐test. C: bars represent mean ± SEM Mann–Whitney U test. Elements created with Figdraw.com .

Article Snippet: The antibodies employed in the study included anti‐SIRT1 (CST, Cat. #D739); anti‐KAP1 (ABclonal, Cat. #A19568), anti‐Lamin B1 (ABclonal, Cat. #A11495), anti‐p21 (ABclonal, Cat. #A19094), anti‐p16 (ABclonal, Cat. #A23882), anti‐TBK1 (CST, Cat. #3504), anti‐Phospho‐TBK1 (CST, Cat. #5483), anti‐STING (ABclonal, Cat. #A21051), anti‐Phospho‐STING (ABclonal, Cat. #AP1369), anti‐H3K9me3 (CST, Cat. #5326), anti‐Flag (ABclonal, Cat. #AE005), anti‐GFP (Abcam, Cat. #ab290), anti‐HA tag (CST, Cat. #3724), anti‐γH2AX (S139) (CST, Cat. #9718), anti‐Tubulin (ABclonal, Cat. #AC007), anti‐rabbit IgG‐HRP (Bio‐Rad, Cat. #170–6515), anti‐rabbit IgG H&L DyLight 488 (Abcam, Cat. #ab96899) and anti‐mouse IgG H&L DyLight 594 (Abcam, Cat. #ab96881).

Techniques: Control, Knockdown, Western Blot, Expressing, Transfection, Plasmid Preparation, Over Expression, Irradiation, Staining, Two Tailed Test, MANN-WHITNEY

Journal: Molecular Biology Reports

Article Title: Icaritin improves pregnancy outcomes in endometriosis by inhibiting ovarian granulosa cell senescence

doi: 10.1007/s11033-026-12629-6

Figure Lengend Snippet: EMS-FF may induce OXPHOS dysfunction and senescence in KGN cells. ( A ) Comparison of ATP concentrations between CON-FF group and EMS-FF group ( n = 3/group). ( B ) Comparison of OCR between CON-FF group and EMS-FF group ( n = 3/group). ( C ) Comparison of basal respiration, maximal respiration, proton leakage, and ATP-coupled respiration levels between CON-FF group and EMS-FF group ( n = 3/group). ( D ) Comparison of ROS levels between CON-FF group and EMS-FF group ( n = 3/group). Scale bar = 100 μm. ( E ) Quantification of ROS levels in (D). ( F ) Quantitative comparison of SA-β-gal-positive cells between CON-FF group and EMS-FF group ( n = 3/group). Scale bar = 100 μm. ( G ) Quantification of SA-β-gal-positive cells in (F). ( H ) Comparison of cell cycle distributions between CON-FF group and EMS-FF group ( n = 3/group). ( I ) Quantification of cell cycle distributions in (H). ( J ) Comparison of P21 mRNA expression levels between CON-FF group and EMS-FF group ( n = 3/group). ( K ) Comparison of P53 mRNA expression levels between CON-FF group and EMS-FF group ( n = 3/group). ( L ) Comparison of P21 protein expression levels between CON-FF group and EMS-FF group ( n = 3/group). ( M ) Comparison of P53 protein expression levels between CON-FF group and EMS-FF group ( n = 3/group). ( N ) Quantification of P21 protein expression levels in (L). ( O ) Quantification of P53 protein expression levels in (M). All data are expressed as Mean ± SD and were analysed by Unpaired Student’s t test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns, not significant

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibodies: anti-NDUFS8 (Abcam, UK; Cat# ab170936), anti-p21 (ABclonal, China; Cat# A19094) and anti-p53 (Proteintech, China; Cat# 60283-2-lg).

Techniques: Comparison, Expressing

Journal: Molecular Biology Reports

Article Title: Icaritin improves pregnancy outcomes in endometriosis by inhibiting ovarian granulosa cell senescence

doi: 10.1007/s11033-026-12629-6

Figure Lengend Snippet: NDUFS8 knockdown may induce senescence in KGN cells via ROS-dependent pathway. ( A ) Comparison of ATP concentration between NC and Sh-NDUFS8 groups ( n = 3/group). ( B ) Comparison of OCR between NC and Sh-NDUFS8 groups ( n = 3/group). ( C ) Comparison of basal respiration, maximal respiration, proton leakage, and ATP-linked respiration between NC and Sh-NDUFS8 groups ( n = 3/group). ( D ) Comparison of ROS levels among NC, Sh-NDUFS8, NC+CoCl₂, and Sh-NDUFS8 + NAC groups ( n = 3/group). Scale bar = 100 μm. ( E ) Quantification of ROS levels in (D). ( F ) Quantitative comparison of SA-β-gal-positive cells in NC, Sh-NDUFS8, NC+CoCl₂, and Sh-NDUFS8 + NAC groups ( n = 3/group). Scale bar = 100 μm. ( G ) Quantification of SA-β-gal-positive cells in (F). ( H ) Comparison of cell cycle distributions among NC, Sh-NDUFS8, NC+CoCl₂, and Sh-NDUFS8 + NAC groups ( n = 3/group). ( I ) Quantification of cell cycle distributions in (H). ( J ) Comparison of P21 mRNA expression levels among NC, Sh-NDUFS8, NC+CoCl₂, and Sh-NDUFS8 + NAC groups ( n = 3/group). ( K ) Comparison of P53 mRNA expression levels among NC, Sh-NDUFS8, NC+CoCl₂, and Sh-NDUFS8 + NAC groups ( n = 3/group). ( L ) Comparison of P21 protein expression levels in among NC, Sh-NDUFS8, NC+CoCl₂, and Sh-NDUFS8 + NAC groups ( n = 3/group). ( M ) Comparison of P53 protein expression levels among NC, Sh-NDUFS8, NC+CoCl₂, and Sh-NDUFS8 + NAC groups ( n = 3/group). ( N ) Quantification of P21 protein expression levels in (L). ( O ) Quantification of P53 protein expression levels in (M). All data are expressed as Mean ± SD and were analysed by Unpaired Student’s t test (between two groups) or One-way ANOVA(between four groups). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns, not significant

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibodies: anti-NDUFS8 (Abcam, UK; Cat# ab170936), anti-p21 (ABclonal, China; Cat# A19094) and anti-p53 (Proteintech, China; Cat# 60283-2-lg).

Techniques: Knockdown, Comparison, Concentration Assay, Expressing

Journal: Molecular Biology Reports

Article Title: Icaritin improves pregnancy outcomes in endometriosis by inhibiting ovarian granulosa cell senescence

doi: 10.1007/s11033-026-12629-6

Figure Lengend Snippet: Ica may inhibit KGN cell senescence by regulating NDUFS8/ROS/P53/P21 axis. ( A ) Comparison of NDUFS8 mRNA expression levels among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). ( B ) Comparison of NDUFS8 protein expression levels among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). ( C ) Quantification of NDUFS8 protein expression levels in (B). ( D ) Comparison of ROS levels among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). Scale bar = 100 μm. ( E ) Quantification of ROS levels in (D). ( F ) Quantitative comparison of SA-β-gal-positive cells among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). Scale bar = 100 μm. ( G ) Quantification of SA-β-gal-positive cells in (F). ( H ) Comparison of cell cycle distributions among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). ( I ) Quantification of cell cycle distributions in (H). ( J ) Comparison of P21 mRNA expression levels among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). ( K ) Comparison of P53 mRNA expression levels among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). ( L ) Comparison of P53 protein expression levels among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). ( M ) Comparison of P21 protein expression levels among CON-FF, EMS-FF and EMS-FF + Ica groups ( n = 3/group). ( N ) Quantification of P21 protein expression levels in (M). ( O ) Quantification of P53 protein expression levels in (L). All data are expressed as Mean ± SD and were analysed by Unpaired Student’s t test (between two groups) or One-way ANOVA (between three groups). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns, not significant

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibodies: anti-NDUFS8 (Abcam, UK; Cat# ab170936), anti-p21 (ABclonal, China; Cat# A19094) and anti-p53 (Proteintech, China; Cat# 60283-2-lg).

Techniques: Comparison, Expressing

Journal: Molecular Biology Reports

Article Title: Icaritin improves pregnancy outcomes in endometriosis by inhibiting ovarian granulosa cell senescence

doi: 10.1007/s11033-026-12629-6

Figure Lengend Snippet: Ica may inhibit GC senescence and improve ovarian function in EMS mice by regulating NDUFS8/ROS/P53/P21 axis. ( A ) H&E staining of ovarian tissues from CON, EMS, Ica, and Die groups ( n = 6/group). Red arrows indicate atretic follicles. Original image scale bar = 200 μm. Enlarged image scale bar = 50 μm. ( B ) Quantification of atretic follicles in (A). ( C ) Comparison of serum AMH levels among CON, EMS, Ica, and Die groups ( n = 6/group). ( D ) Comparison of serum FSH levels among CON, EMS, Ica, and Die groups ( n = 6/group). ( E ) Comparison of serum LH levels among CON, EMS, Ica, and Die groups ( n = 6/group). ( F ) IHC NDUFS8 expression levels in ovarian tissues from CON, EMS, Ica, and Die groups ( n = 6/group). Original image scale bar = 200 μm. Enlarged image scale bar = 50 μm. ( G ) Quantification of NDUFS8 expression levels in (F). ( H ) IHC P21 expression levels in ovarian tissues from CON, EMS, Ica, and Die groups ( n = 6/group). Original image scale bar = 200 μm. Enlarged image scale bar = 50 μm. ( I ) Quantification of P21 expression levels in (H). ( J ) IHC P53 expression levels in ovarian tissues from CON, EMS, Ica, and Die groups ( n = 6/group). Original image scale bar = 200 μm. Enlarged image scale bar = 50 μm. ( K ) Quantification of P53 expression levels in (J). ( L ) Comparison of ROS levels in mouse primary GCs among CON, EMS, Ica, and Die groups ( n = 3/group). Scale bar = 100 μm. ( M ) Quantification of ROS levels in (L). ( N ) Quantitative comparison of SA-β-gal-positive cells in mouse primary GCs among CON, EMS, Ica, and Die groups ( n = 3/group). Scale bar = 100 μm. ( O ) Quantification of SA-β-gal-positive cells in (N). ( P ) Comparison of cell cycle distributions in mouse primary GCs among CON, EMS, Ica, and Die groups ( n = 3/group). ( Q ) Quantification of cell cycle distributions in (P). All data are expressed as Mean ± SD and were analysed by Unpaired Student’s t test (between two groups) or One-way ANOVA (between four groups). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, and ns, not significant

Article Snippet: Sections were incubated overnight at 4°C with the following primary antibodies: anti-NDUFS8 (Abcam, UK; Cat# ab170936), anti-p21 (ABclonal, China; Cat# A19094) and anti-p53 (Proteintech, China; Cat# 60283-2-lg).

Techniques: Staining, Comparison, Expressing

Journal: Aging Cell

Article Title: Diosgenin Alleviates Age‐Related Sarcopenia by Promoting Satellite Cell Proliferation and Myogenic Differentiation via Activation of the SIRT1 / PGC ‐1α Signaling Pathway

doi: 10.1111/acel.70651

Figure Lengend Snippet: Diosgenin promoted proliferation and myogenic differentiation of senescent satellite cells. (A) Representative images of Pax7/Ki67 double IF staining in skeletal muscle tissues, with quantitative analysis. Green: Pax7 (satellite cell marker); red: Ki67 (proliferation marker); blue: DAPI (nuclei); yellow: Pax7 + /Ki67 + double‐positive cells. Scale bar = 20 μm. (B) Cytotoxicity of diosgenin in differentiated myotubes (determination of safe concentration range, n = 3). (C) Proliferative activity of diosgenin in undifferentiated C2C12 myoblasts in the absence (left panel) or presence (right panel) of 100 μM D‐gal ( n = 3). (D) Representative WB images and quantitative analysis of the myogenic differentiation markers Myf5, Pax7, and MyHC II in mouse skeletal muscle. GAPDH was used as the internal control. (E) Representative WB images and quantitative analysis of the myogenic differentiation markers Myf5, Pax7, and MyHC II in C2C12 cells. GAPDH was used as the internal control. Data are presented as mean ± SEM. * p ‐value < 0.05, ** p ‐value < 0.01, *** p ‐value < 0.001 versus the YC group; # p ‐value < 0.05, ## p ‐value < 0.01, ### p ‐value < 0.001 versus the OC group. Statistical analysis was performed using one‐way ANOVA followed by Tukey's multiple comparisons test. All experiments were performed with at least three independent replicates.

Article Snippet: Membranes were blocked with 5% skim milk at room temperature for 1 h and incubated with the following primary antibodies at 4°C overnight: SIRT1 (1:1000; Affinity Biosciences, Liyang, China; Cat. #DF6033), PGC‐1α (1:1000; ABclonal, Wuhan, China; Cat. #AP20542C), p53 (1:1000; ABclonal, Wuhan, China; Cat. #A21630), p21 (1:1000; ABclonal, Wuhan, China; Cat. #A1483), Myf5 (1:1000; ABclonal, Wuhan, China; Cat. #A116227), MyHC II (1:1000; ABclonal, Wuhan, China; Cat. #A15293), Pax7 (1:1000; ABclonal, Wuhan, China; Cat. #A7335), and GAPDH (1:20,000; ABclonal, Wuhan, China; Cat. #AC033).

Techniques: Cell Characterization, Staining, Marker, Concentration Assay, Activity Assay, Control

Journal: Aging Cell

Article Title: Diosgenin Alleviates Age‐Related Sarcopenia by Promoting Satellite Cell Proliferation and Myogenic Differentiation via Activation of the SIRT1 / PGC ‐1α Signaling Pathway

doi: 10.1111/acel.70651

Figure Lengend Snippet: Diosgenin regulated SIRT1 to ameliorate age‐related sarcopenia. (A) Representative WB images and quantitative analysis of SIRT1 in C2C12 cells. (B) Quantitative analysis of PGC‐1α protein expression and PGC‐1α acetylation levels in C2C12 cells. (C) mRNA levels of Cpt1b in C2C12 cells. (D) mRNA levels of Metrnl and Igf1 in C2C12 cells. (E) Protein concentrations of METRNL and IGF‐1 in C2C12 cells. (F) Representative WB images and quantitative analysis of the myogenic differentiation markers Myf5, Pax7, and MyHC II in C2C12 cells. (G) Representative WB images and quantitative analysis of the senescence markers p53 and p21 in C2C12 cells. (H) Representative SA‐β‐gal staining images of C2C12 cells at 100× and 400× magnification, with quantitative analysis. (I) Molecular docking results of diosgenin with the SIRT1 protein. (J) Schematic diagram of the interactions between diosgenin and key residues of SIRT1. (K) Representative WB images reflecting the thermal stability of SIRT1 protein under different temperature gradients and the corresponding thermal stability curve; the y‐axis represents the remaining SIRT1 protein level relative to 37°C. Data are presented as mean ± SEM. * p‐ value < 0.05, ** p ‐value < 0.01, *** p ‐value < 0.001 versus the YC group; # p ‐value < 0.05, ## p ‐value < 0.01, ### p ‐value < 0.001 versus the OC group. Statistical analysis was performed using one‐way ANOVA followed by Tukey's multiple comparisons test. All experiments were performed with at least three independent replicates.

Article Snippet: Membranes were blocked with 5% skim milk at room temperature for 1 h and incubated with the following primary antibodies at 4°C overnight: SIRT1 (1:1000; Affinity Biosciences, Liyang, China; Cat. #DF6033), PGC‐1α (1:1000; ABclonal, Wuhan, China; Cat. #AP20542C), p53 (1:1000; ABclonal, Wuhan, China; Cat. #A21630), p21 (1:1000; ABclonal, Wuhan, China; Cat. #A1483), Myf5 (1:1000; ABclonal, Wuhan, China; Cat. #A116227), MyHC II (1:1000; ABclonal, Wuhan, China; Cat. #A15293), Pax7 (1:1000; ABclonal, Wuhan, China; Cat. #A7335), and GAPDH (1:20,000; ABclonal, Wuhan, China; Cat. #AC033).

Techniques: Expressing, Cell Characterization, Staining