Journal: bioRxiv

Article Title: Senescence-inhibitory Δ133p53α counteracts accelerated ageing and mortality

doi: 10.64898/2025.12.31.697195

Figure Lengend Snippet: Western blot analyses of lamin A/C and progerin, full-length p53, Δ133p53α, and p21 Waf1/Cip1 were performed in four 15-week-old Δ133p53α-expressing Group-1 mice ( CAG-133 Tam/+ ; Cre Tg/+ ; Lmna G609G/+ ), along with four each of age-matched, non-expressing control Group-2 ( CAG-133 LSL/+ ; Cre Tg/+ ; Lmna G609G/+ ) and Group-4 mice ( CAG-133 +/+ ; Cre +/+ ; Lmna G609G/+ ), as well as two age-matched wild-type mice ( CAG-133 +/+ ; Cre +/+ ; Lmna +/+ ). Results from skin ( a ), skeletal muscle ( b ), kidney ( c ), spleen ( d ), and lung ( e ) are presented. An inter-blot control (liver from a Group-1 mouse) was included in all blots. F, female; M, male. GAPDH was a loading control and used for normalization of p21 Waf1/Cip1 , progerin, and full-length p53 expression levels. Quantitative data summaries of p21 Waf1/Cip1 , progerin, and full-length p53 in Group-1, -2 and -4 mice are shown as relative values to Group-2 mice (mean ± s.d. from n = 4; open circles indicate two females, and closed circles indicate two males). P values were determined by Welch’s t -test. Two wild-type mice were used only as references and not for statistical comparisons.

Article Snippet: Primary antibodies used were as follows: anti-p53 antibodies SAPU (sheep polyclonal) and DO-11 (mouse monoclonal; Bio-Rad, MCA1704) ; anti-p21 Waf1/Cip1 (mouse monoclonal; Santa Cruz Biotechnology, sc-6246, clone F-5); anti-lamin A/C (mouse monoclonal; Santa Cruz Biotechnology, sc-376248); anti-GAPDH (mouse monoclonal; Santa Cruz Biotechnology, sc-166574); and anti-β-actin (mouse monoclonal; Thermo Fisher Scientific, MA1-91399).

Techniques: Western Blot, Expressing, Control

Journal: bioRxiv

Article Title: Senescence-inhibitory Δ133p53α counteracts accelerated ageing and mortality

doi: 10.64898/2025.12.31.697195

Figure Lengend Snippet: a,b , Top Hallmark pathways identified by Gene set enrichment analysis (GSEA). The bulk RNA-seq data were obtained from the heart ( a ) and kidney ( b ) in 9-10-month-old Group-1 and Group-3 mice (n = 5 each). Pathways are ranked by normalized enrichment score. False discovery rate < 0.10. c-g, Enrichment plots illustrate significant downregulation of the p53 pathway ( c,d ) and upregulation of the oxidative phosphorylation ( e,f ) in both heart and kidney, as well as upregulation of the glycolysis in the kidney ( g ). Enrichment plots depict running enrichment scores (ES) and the distribution of genes within each Hallmark gene set. All leading edge genes in each pathway are listed in Extended Data Table 2. h,i, qRT-PCR assays of mRNA expression of genes in the oxidative phosphorylation pathway (Ndufs6, Ndufc2, Uqcrq, Hsd17b10, and Gpx4) and an antioxidant gene Prdx1 in the heart ( h ) and kidney ( i ) of 9-10-month-old Group-1 and Group-3 mice (mean ± s.d. from n = 5, each with technical triplicate; open circles, females; closed circles, males). P values were calculated by Welch’s t -test.

Article Snippet: Primary antibodies used were as follows: anti-p53 antibodies SAPU (sheep polyclonal) and DO-11 (mouse monoclonal; Bio-Rad, MCA1704) ; anti-p21 Waf1/Cip1 (mouse monoclonal; Santa Cruz Biotechnology, sc-6246, clone F-5); anti-lamin A/C (mouse monoclonal; Santa Cruz Biotechnology, sc-376248); anti-GAPDH (mouse monoclonal; Santa Cruz Biotechnology, sc-166574); and anti-β-actin (mouse monoclonal; Thermo Fisher Scientific, MA1-91399).

Techniques: RNA Sequencing, Phospho-proteomics, Quantitative RT-PCR, Expressing

Journal: American Journal of Cancer Research

Article Title: Gentian violet induces apoptosis and ferroptosis via modulating p53 and MDM2 in hepatocellular carcinoma

doi:

Figure Lengend Snippet: GV simultaneously increased the expression levels of MDM2 and p53. A. SK-HEP-1 and SMMC-7721 cells were treated without or with GV at the concentration of 50, 100 nM or 150, 300 nM respectively, for 48 h. Immunoblot was carried out with the indicated antibodies. B. qRT-qPCR results showing the relative fold changes of MDM2 and p53 mRNA expression at the same treatment condition. C. Representative images of the H&E staining, and the p53 and MDM2 immunohistochemical (IHC) staining of the xenograft tumor tissues. Data were presented as mean ± SD. **p<0.01.

Article Snippet: Knockdown of MDM2, p53 and Hep27 was performed using siRNA technology. siRNA oligos targeting MDM2 (sc-29394), p53 (sc-29435), Hep27 (sc-92153) or non-specific siRNAs were purchased from Santa Cruz Biotechnology (Dallas, TX). siRNAs were transfected into cells by electroporation using Nucleofector Device and the Nucleofector Kit L (Lonza, Basel, Switzerland).

Techniques: Expressing, Concentration Assay, Western Blot, Staining, Immunohistochemical staining, Immunohistochemistry

Journal: American Journal of Cancer Research

Article Title: Gentian violet induces apoptosis and ferroptosis via modulating p53 and MDM2 in hepatocellular carcinoma

doi:

Figure Lengend Snippet: The high expression of MDM2 and p53 was dependent on Hep27. Western blot detection of MDM2, p53 and Hep27 expression in SK-HEP-1 and SMMC-7721 cells at the indicated treatment conditions for 48 h. From left to right for each cell line: DMSO, GV (100 nM for SK-HEP-1 and 300 nM for SMMC-7721), GV in cells transfected with Hep27 siRNA, GV co-treated with Fer-1 (2 μM).

Article Snippet: Knockdown of MDM2, p53 and Hep27 was performed using siRNA technology. siRNA oligos targeting MDM2 (sc-29394), p53 (sc-29435), Hep27 (sc-92153) or non-specific siRNAs were purchased from Santa Cruz Biotechnology (Dallas, TX). siRNAs were transfected into cells by electroporation using Nucleofector Device and the Nucleofector Kit L (Lonza, Basel, Switzerland).

Techniques: Expressing, Western Blot, Transfection

Journal: American Journal of Cancer Research

Article Title: Gentian violet induces apoptosis and ferroptosis via modulating p53 and MDM2 in hepatocellular carcinoma

doi:

Figure Lengend Snippet: The proposed molecular mechanisms of GV-induced killing of HCC. When NOX is inhibited by GV, Hep27 is increased, which leads to the blockade of the ubiquitination and degradation of p53 by MDM2. Consequently, the levels of p53 and MDM2 are increased. GV also directly increases p53 level. Increased p53 and MDM2 levels trigger ferroptosis and mitochondrial apoptosis (through the activation of caspase 8) by a coordinated ROS cross point and downstream signaling cascades. On the other hand, GV also triggers death receptor apoptosis (through the activation of caspase 9) by upregulating death receptors DR4/5 and ligands FAS-ligand and TRAIL.

Article Snippet: Knockdown of MDM2, p53 and Hep27 was performed using siRNA technology. siRNA oligos targeting MDM2 (sc-29394), p53 (sc-29435), Hep27 (sc-92153) or non-specific siRNAs were purchased from Santa Cruz Biotechnology (Dallas, TX). siRNAs were transfected into cells by electroporation using Nucleofector Device and the Nucleofector Kit L (Lonza, Basel, Switzerland).

Techniques: Ubiquitin Proteomics, Activation Assay