phosphorylated p53  (Cell Signaling Technology Inc)

Journal: Oncology Research

Article Title: miR-100-5p Enhances Cell Cycle-Mediated Chemoresistance by Modulating the CTDSPL/pRB/E2F1 Signaling Pathway in Oxaliplatin-Resistant Colorectal Cancer Cells

doi: 10.32604/or.2026.073080

Figure Lengend Snippet: Lower CTDSPL expression is significantly associated with cell cycle alteration in OXR-LoVo cells. ( A ) Western blot analysis showing the expression levels of CTDSPL, pRB, RB and E2F1; ( B , C ) the cell cycle phase regulatory proteins (Cdc25a, cyclin E, cyclin A, cyclin D1, PCNA, Cdk4, Cdk6 and cyclin B1); ( D ) cell cycle inhibitory proteins (p-p53, p27, p15/16, p19) in parental and oxaliplatin-resistant LoVo cells. GAPDH was used as endogenous control. Quantification of data was carried out by ImageJ software 1.50 and is presented as the mean ± SD ( n = 3). Significance ascribed as ** p < 0.01, *** p < 0.001 vs. oxaliplatin-resistant LoVo cells.

Article Snippet: CTDSPL (NBP1-91814) (Novus Biologicals Co., Centennial, CO, USA); E2F1 (#3742), phosphorylated RB (#9308), and RB (#9309), p53 (#2524), phosphorylated-p53 (#9284) (Cell Signaling Technology, Inc. Beverly, MA, USA); cyclin E1 (ab33911), cyclin A1 (ab270940) (Abcam, Cambridge, UK); and E-cadherin (sc-8426), Vimentin (sc-32322), α-Tubulin (sc-5286), β-Catenin (sc-7963), Cdk4 (sc-23896), Cdk6 (sc-7961), Cdc25a (sc-7389), cyclin D1 (sc-246), cyclin B1 (sc-752), cyclin A1, PCNA (sc-56), ERK1/2 (sc-514032), phosphorylated-ERK1/2 (sc-7383), AKT (sc-5298), phosphorylated-AKT (sc-514032), p19 (sc-166774), p27 (sc-1641), p15/16 (sc-377412), Ki67 (sc-23900) and GAPDH (sc-32233) (Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA).

Techniques: Expressing, Western Blot, Control, Software

Journal: International Journal of Molecular Sciences

Article Title: Dysfunction of the Autophagy System and MDM2–p53 Axis Leads to the Accumulation of Amyloidogenic Proteins in Angelman Syndrome Models

doi: 10.3390/ijms262211032

Figure Lengend Snippet: MDM2–p53 axis in the AS neuronal cell model. ( A , B ) Cell lysates from control and UBE3A − SH-SY5Y cells were used for western blotting analysis using specific antibodies to MDM2. A ‘stain-free protein normalization’ method was used for the normalization of bands to total protein in blots, eliminating the need for housekeeping proteins. ( C ) Cell lysates from control and UBE3A − SH-SY5Y cells were used to detect MDM2-p53 complex by a specific immunoezymatic assay, as reported in the . Data are expressed as percentage versus control cells and are mean ± SEM from four independent experiments. Comparisons between control and UBE3A − were performed using an unpaired two-tailed Student’s t -test; *** p < 0.001 control vs. UBE3A − .

Article Snippet: The following primary antibodies were employed: anti-MDM2 (sc-5304, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500); p-62 (SQSTM1/p62, #5114, Cell Signaling Technology, Danvers, MA, USA, diluted 1:1000); LC3β (sc-271625, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500); total p53 (sc-126, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500); Phosphorylated p-p53 (sc-101762, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500).

Techniques: Control, Western Blot, Staining, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Dysfunction of the Autophagy System and MDM2–p53 Axis Leads to the Accumulation of Amyloidogenic Proteins in Angelman Syndrome Models

doi: 10.3390/ijms262211032

Figure Lengend Snippet: p53 expression and activity in SH-SY5Y AS cellular model. ( A – D ) Cell lysates from SH-SY5Y (control and UBE3A − cells), were used for western blotting analysis using specific antibodies to total p53 ( A , B ) and phosphorylated p53 ( C , D ). A ‘stain-free protein normalization’ method was used for the normalization of bands to total protein in blots, eliminating the need for housekeeping proteins. Data are expressed as percentage versus control cells and are mean ± SEM of at least three independent experiments Comparisons between control and UB3A − were performed using an unpaired two-tailed Student’s t -test; * p < 0.05, ** p < 0.01, UBE3A − vs. control. ( E ) SHSY-5Y (control and UBE3A − ) cells were collected for RNA extraction and retro transcription to cDNA. A real-time PCR analysis was performed to monitor mRNA expression of p53-traget genes (i.e., Bax, p21, and MDM2). Data are expressed as fold change relative to Control cells (set to 1) and represent mean ± SEM of four independent experiments. Comparisons between Control and UBE3A − were performed using an unpaired two-tailed Student’s t -test; *** p < 0.001.

Article Snippet: The following primary antibodies were employed: anti-MDM2 (sc-5304, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500); p-62 (SQSTM1/p62, #5114, Cell Signaling Technology, Danvers, MA, USA, diluted 1:1000); LC3β (sc-271625, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500); total p53 (sc-126, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500); Phosphorylated p-p53 (sc-101762, Santa Cruz Biotechnology, Santa Cruz, CA, USA, diluted 1:500).

Techniques: Expressing, Activity Assay, Control, Western Blot, Staining, Two Tailed Test, RNA Extraction, Real-time Polymerase Chain Reaction