Journal: Nature Communications

Article Title: Phosphorylation of PA28γ by CK2 kinase facilitates HNSCC tumor formation and progression

doi: 10.1038/s41467-025-67131-7

Figure Lengend Snippet: a IB analysis of WCLs derived from HEK293T cells transfected with His-E4F1 and increasing doses of Flag-PA28γ. b Quantitative real-time PCR analysis of the indicated genes in HEK293T cells transfected with EV and increasing doses of Flag-PA28γ. Endo., endogenous. c HSC-3 cells stably expressing EV or Flag-PA28γ were treated with 100 μgml −1 CHX for the indicated time before harvesting. Quantification of E4F1 levels relative to tubulin levels is shown. d , e HEK293T cells stably expressing EV or Flag-PA28γ were transfected with Myc-E4F1 truncations, and treated with 100 μgml −1 CHX for the indicated time before harvesting. Quantification of Myc-E4F1 levels relative to tubulin levels is shown. f HEK293T cells transfected with the indicated plasmids were treated without or with 10 μM MG132 for 6 h before harvesting, and WCLs were collected for IB analysis. g HEK293T cells stably expressing shRNA against endogenous PA28γ were transfected with the indicated plasmids, and WCLs were collected for IB analysis. h Purified PA28γ, E4F1 and 20S proteasome in the absence or presence of 100 nM proteasome inhibitor epoxomicin (Epox) were incubated as indicated for 45 min, followed by IB analysis. i Purified PA28γ WT, PA28γ-T23A, E4F1, p21 (as a positive control for E4F1) and 20S proteasome were incubated as indicated for 45 min, followed by IB analysis. Data in ( b – e ) represent the mean ± SD of three biological replicates; statistical significance was assessed by two-sided unpaired t -test. Source data are provided as a Source Data file.

Article Snippet: Recombinant human E4F1 protein (H00001877-P01) and recombinant human p21/CDKN1A protein (NBP2-22976) were purchased from Novus Biologicals.

Techniques: Derivative Assay, Transfection, Real-time Polymerase Chain Reaction, Stable Transfection, Expressing, shRNA, Purification, Incubation, Positive Control

Journal: Journal of Veterinary Research

Article Title: Immunoreactivity of p21, MMP-1 and CB2 receptor proteins in cutaneous canine mast cell tumours: an association with the three-tier grading system

doi: 10.2478/jvetres-2023-0066

Figure Lengend Snippet: Immunohistochemistry with primary anti-p21 antibody with positive reaction visible as brown staining. A – weak nuclear reaction in a grade 1 mast cell tumour (MCT); B – moderate nuclear reaction in a grade 2 MCT; C – strong nuclear reaction in a grade 3 MCT

Article Snippet: The primary antibodies used in this study, all canine-specific, were as follows: anti-CB2R (Cat. No. TA317640; OriGene Technologies, Rockville, MD, USA), anti-p21 (Cat. No. M7202; clone SX118, Agilent Dako, Santa Clara, CA, USA) and anti-MMP-1 (Cat. No. ABIN2777120; antibodies-online, Aachen, Germany), diluted 1 : 200, 1 : 50 and 1 : 500, respectively.

Techniques: Immunohistochemistry, Staining

Journal: Journal of Veterinary Research

Article Title: Immunoreactivity of p21, MMP-1 and CB2 receptor proteins in cutaneous canine mast cell tumours: an association with the three-tier grading system

doi: 10.2478/jvetres-2023-0066

Figure Lengend Snippet: Frequency of immunoreactivity for p21 protein in grade 1, 2 and 3 mast cell tumours (MCTs). * – P-value < 0.05; **_ P-value < 0.01; *** P-value < 0.001 compared to grade 1 MCTs; (−) – no reaction; (+) – weak reaction; (++) – moderate reaction; (+++) – strong reaction

Article Snippet: The primary antibodies used in this study, all canine-specific, were as follows: anti-CB2R (Cat. No. TA317640; OriGene Technologies, Rockville, MD, USA), anti-p21 (Cat. No. M7202; clone SX118, Agilent Dako, Santa Clara, CA, USA) and anti-MMP-1 (Cat. No. ABIN2777120; antibodies-online, Aachen, Germany), diluted 1 : 200, 1 : 50 and 1 : 500, respectively.

Techniques:

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: (A-D) The analysis of male 19 months WT or Gzmk -/- mice (male). (A) Staining of SA-β-gal in liver and hippocampus. (B) H&E staining in liver, lung and kidney. (C) SA-β-gal staining for WAT from indicated mice. (D) Western blot analysis of the expression of p16, p21 and p53 in liver. (E) The bioluminescence imaging of indicated mice (15 months, female) by injecting luciferase substrates. (F-G) CD8 T cells (4 x10 6 ) from old or old Gzmk -/- mice were adoptively transferred into young p16 Ink4a -luciferase reporter mice, one month later, the luciferase activity was measured (F), and the expression of the p16 -driven luciferase reporter in indicated tissues were measured by RT-qPCR.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Staining, Western Blot, Expressing, Imaging, Luciferase, Activity Assay, Quantitative RT-PCR

Journal: bioRxiv

Article Title: A feed-forward loop between niche adenosine and Gzmk⁺ CD8 T cells propagates systemic inflammaging

doi: 10.64898/2026.03.18.712515

Figure Lengend Snippet: Old mice (19 months) were administrated with SCH or PPACK for one-month. The level of IL-6 and TNFα (A), ALT and AST (B) in plasma from indicated mice were determined by ELSIA. (C) Staining of SA-β-gal in liver. (D) H&E staining of lungs from indicated mice. (E) Staining of SA-β-gal in hippocampus from indicated mice. (F-G) Western blot analysis of P16, P21 and P53 at liver and brain from indicated mice.

Article Snippet: Primary antibodies used in this study: Mouse anti-CDKN2A/p16 (Santa Cruz Biotechnology, cat# sc-1661, 1:1000) Mouse anti-CDKN1A p21 (Santa Cruz Biotechnology, cat# sc-6246, 1:1000) Mouse anti-p53 (Santa Cruz Biotechnology, cat# sc-98, 1:1000) Mouse anti-GAPDH (Absin, abs830030,1:10000) Rabbit anti-γH2A.X (Cell Signaling Technology, cat# 9718S, 1:1000)

Techniques: Clinical Proteomics, Staining, Western Blot

Journal: Oncogene

Article Title: Myostatin inhibits rhabdomyosarcoma cell proliferation through an Rb-independent pathway.

doi: 10.1038/sj.onc.1207144

Figure Lengend Snippet: Figure 5 Myostatin does not alter the levels of the CKIs p21, p15, p16 or p27 in RMS cells. Western blots showing the levels of p21, p15, p16 and p27 protein in RD cells cultured with ( þ ) or without () myostatin for 24 and 48 h. p21, p15, p16 and p27 immunoreactive proteins were detected using their respective antibodies. Tubulin protein levels, detected by antitubulin anti- bodies, are included to show even loadings

Article Snippet: The following primary antibodies were used for immunoblotting; myostatin: 1 : 2000 dilution of rabbit polyclonal antimyostatin antibody (Thomas et al., 2000); p15, 1 : 400 dilution of purified rabbit polyclonal anti-p15 antibody (sc-613; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); p16, 1 : 400 dilution of purified rabbit polyclonal antip16 antibody (sc-1207; Santa Cruz Biotechnology Inc.); p21, 1 : 400 dilution of purified mouse monoclonal anti-p21 antibody (SX118; PharMingen, CA, USA); p27, 1 : 400 dilution of purified mouse monoclonal anti-p27 antibody (sc-1641; Santa Cruz Biotechnology Inc.); Cdk2, 1 : 400 dilution of purified mouse monoclonal anti-Cdk2 antibody (sc-2648; Santa Cruz Biotechnology Inc.); Cdk4, 1 : 400 dilution of purified rabbit polyclonal anti-Cdk4 antibody (sc-601; Santa Cruz Biotechnology Inc.); cyclin-D1, 1 : 400 dilution of purified mouse monoclonal anticyclin-D1 antibody (sc-8396; Santa Cruz Biotechnology Inc.); cyclin-E, 1 : 400 dilution of purified rabbit polyclonal anticyclin-E antibody (sc-481; Santa Cruz Biotechnology Inc.); a-tubulin, 1 : 3000 dilution of purified mouse monoclonal anti-a-tubulin antibody (DM 1A; Sigma); Rb, 1 : 200 dilution of purified mouse monoclonal anti-pRb antibody (G3-245; PharMingen); E2F1, 1 : 250 dilution of purified mouse monoclonal anti-E2F1 antibody (KH95; PharMingen); NPAT, 1 : 250 dilution of purified mouse monoclonal antiNPAT antibody (C27; Transduction Laboratories) or 1 : 2000 rabbit polyclonal anti-NPAT antibody (Zhao et al., 2000).

Techniques: Western Blot, Cell Culture

Journal: Biochemical and biophysical research communications

Article Title: Loss of DNA replication fork protection by TIMELESS degradation supports oncogene-induced senescence

doi: 10.1016/j.bbrc.2025.152203

Figure Lengend Snippet: (A) Schematic of PARP1-mediated TIM PARylation and PAR-dependent proteolysis. (B-D) WB of U2OS parental or PARP1 KO clones transduced with pBABE-HRAS G12V (vs. EV) for 24 h. Where indicated, 10 μM olaparib (ola) or talazoparib (tal) was co-treated for 20 h before harvest. (E) WB of IMR90 ER:HRAS G12V induced by 100 nM 4-OHT in the presence or absence of 10 μM olaparib. (F, G) WB of BJ-5ta ER:HRAS G12V induced by 4-OHT. For (G), cells were co-treated with indicated inhibitors for 16 h. (H) Anti-Flag immunoprecipitation (IP) of Flag-TIM (pcDNA4) in denaturing conditions followed by anti-pADPr WB in U2OS ER:HRAS G12V induced by 4-OHT and treated with indicated inhibitors for 16 h. (I) As (H), but in cells expressing Flag-TIM wild-type (WT) or PBM1/2. (J) A model depicting the signaling cascade that connects oncogenic RAS to PARP1, prompting PAR-dependent proteasomal degradation of TIM.

Article Snippet: Following antibodies were used: TIMELESS (Bethyl, A300–961A), RB (Santa Cruz, sc-102), HRAS (Proteintech, CL488–18295), Poly(ADP-ribose) (R&D, 4336-BPC-100), PCNA (Santa Cruz, sc-56), Cyclin A (Santa Cruz, sc-271682), and anti-Flag M2 affinity gel (Sigma, A2220).

Techniques: Activation Assay, Clone Assay, Transduction, Immunoprecipitation, Expressing