Journal: bioRxiv

Article Title: IRES-mediated translation of Δ160p53 regulates p53 functions and fine-tunes cancer homeostasis

doi: 10.64898/2026.08.21.744132

Figure Lengend Snippet: (A) Luciferase assay of H1299 cells after 48 h of transfection with pcDNA, FL-p53, and Δ160p53, and luciferase constructs (PG13 and pRL-TK); the PG13 construct is a p53 binding site-containing firefly luciferase plasmid, and pRL-TK is used as a transfection control. (B) Real-time PCR analysis of p21 mRNA levels in H1299 cells transfected with pcDNA, FL-p53, and Δ160p53 and luciferase constructs (PG13 and pRL-TK). (C) Western blot validation of FL-p53, Δ160p53, and p21 expression from H1299 cells transfected with pcDNA, FL-p53, and Δ160p53 and luciferase constructs (PG13 and pRL-TK), probed with p21 antibody, p53 polyclonal antibody, and β-actin antibody. (D) Luciferase assay of A549 cells transfected with pcDNA, Δ160p53, and luciferase constructs (PG13 and pRL-TK). (E) Luciferase assay of HCT116+/+ cells transfected with pcDNA, Δ160p53, and luciferase constructs (PG13 and pRL-TK). (F) Luciferase assay of HeLa cells after 48 h of transfection with pcDNA, Δ160p53, and luciferase constructs (PG13 and pRL-TK). (G) Western blot validation of FL-p53 and Δ160p53 from A549 cells transfected with pcDNA, Δ160p53, and luciferase constructs (PG13 and pRL-TK), probed with p53 polyclonal antibody and β-actin antibody. (H) Western blot analysis of FL-p53 and Δ160p53 from HCT116+/+ cells transfected with pcDNA, Δ160p53, and luciferase constructs (PG13 and pRL-TK), probed with p53 polyclonal antibody and β-actin antibody. (I) Western blot validation of FL-p53 and Δ160p53 in HeLa after 48 h of transfection with pcDNA, Δ160p53, and luciferase constructs (PG13 and pRL-TK), probed with p53 polyclonal antibody and β-actin antibody. Error bars indicate standard deviation (SD). All experiments were performed in three biological replicates (n = 3). A two-tailed Student’s t-test determined the criterion for significance (*P ≤ 0.05 or **P ≤ 0.01 or ***P ≤ 0.001).

Article Snippet: The BiP antibody (Abclonal, Cat. No. A4908), rabbit anti-GFP-Tag antibody (Abclonal, Cat. No. AE011), and p21 antibody (Abclonal, Cat. No. A19094) were used to probe Bip, GFP, and p21, respectively.

Techniques: Luciferase, Transfection, Construct, Binding Assay, Plasmid Preparation, Control, Real-time Polymerase Chain Reaction, Western Blot, Biomarker Discovery, Expressing, Standard Deviation, Two Tailed Test

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: Bioactive Materials

Article Title: Gynostemma pentaphyllum -derived extracellular vesicles alleviate skin aging by destabilizing STING

doi: 10.1016/j.bioactmat.2026.03.010

Figure Lengend Snippet: GPEVs improve d UVB irradiation-induced senescence of HEKa cells in vitro. (A) The SA-β-gal staining of UVB-irradiated HEKa (6 mJ/cm 2 ) cells subjected to treatments with a range of GPEVs concentrations (1 × 10 8 , 2 × 10 8 , 4 × 10 8 particles/ml) and NAC (1 mM). Scale bar: 50 μm. (B) The Quantification of SA-β-gal staining in A. (C) Representative images of EdU staining in UVB-irradiated HEKa cells following treatment with NAC and varying concentrations of GPEVs. Scale bar: 50 μm. (D) Quantification of EdU-positive cells in C. (E) The protein levels of p16, p21 and p53 in HEKa cells exposed to UVB irradiation and GPEVs treatment were assessed using Western blot analysis. (F) The quantification of E was performed using Image J software. (G) The relative mRNA levels of the senescence-associated secretory phenotype in irradiated HEKa cells treated with or without GPEVs were measured by RT-qPCR. All the experiments were repeated at least three times. Data are represented as means ± SD. Statistical analysis was performed using one-way ANOVA followed by Bonferroni post hoc test. Not significant (ns), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The primary antibodies used included γ-H2AX and CTCF (1:1,000, Cell Signaling), p21 and STING (diluted 1:1,000, Cell Signaling).

Techniques: Irradiation, In Vitro, Staining, Western Blot, Software, Quantitative RT-PCR

Journal: Bioactive Materials

Article Title: Gynostemma pentaphyllum -derived extracellular vesicles alleviate skin aging by destabilizing STING

doi: 10.1016/j.bioactmat.2026.03.010

Figure Lengend Snippet: The application of GPEVs effectively mitigate d UVB-induced skin aging in Balb/C mice. (A) Representative images of Masson's trichrome staining of mice skin from each group. Collagen fibers are stained blue. Scale bar: 200 μm. (B) Quantitative analysis of collagen fibers in Masson's trichrome stained skin tissue sections by Image J software. (C and D) The levels of p21 in mice skin were detected by immunofluorescent staining and quantified using Image J software. (E) The protein levels of aging-related proteins in skin tissues were determined by Western Blots. (F) The quantification of E was conducted using Image J software. (G) The mRNA levels of senescence-associated secretory phenotype in skin tissues were detected by RT-qPCR. Data are represented as means ± SD. Statistical analysis was performed using One-way ANOVA followed by Bonferroni post hoc test. Not significant (ns), ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

Article Snippet: The primary antibodies used included γ-H2AX and CTCF (1:1,000, Cell Signaling), p21 and STING (diluted 1:1,000, Cell Signaling).

Techniques: Staining, Software, Western Blot, Quantitative RT-PCR