Journal: Experimental and Therapeutic Medicine

Article Title: Purpurogallin carboxylic acid exhibits synergistic effects with 5‑fluorouracil on liver cancer cells in vitro by targeting ABCG2

doi: 10.3892/etm.2024.12564

Figure Lengend Snippet: PCA combined with 5-FU induces the G1 phase cell cycle arrest in liver cancer cells. (A) Flow cytometry was carried out to evaluate the cell cycle distribution in HepG2, Huh7 and Huh1 cells treated with DMSO, 10 µM PCA, 10 µM 5-FU or their combination. Western blot analysis was performed to detect the protein expression levels of CDK4 and CDK6 in (B) HepG2, (C) Huh7 and (D) Huh1 cells treated with DMSO, 10 µM PCA, 10 µM 5-FU or their combination. Reverse transcription- quantitative PCR was performed to detect the mRNA levels of CDK4 and CDK6 in (E) HepG2, (F) Huh7 and (G) Huh1 cells treated with DMSO, 10 µM PCA, 10 µM 5-FU or their combination. * P<0.05 and ** P<0.01. PCA, purpurogallin carboxylic acid; 5-FU, 5-fluorouracil; CDK, cyclin-dependent kinase.

Article Snippet: Following blocking with 8% skim milk powder in TBS-Tween-20 (0.1%) (TBST) for 2 h at 28 ̊C, the membranes were incubated with primary antibodies against cyclin-dependent kinase (CDK) 4 (1:2,000; cat no. 11026-1-AP), CDK6 (1:1,000; cat no. 14052-1-AP), ABCG2 (1:1,000; cat no. 27286-1-AP) and GAPDH (1:50,000; cat no. 60004-1-Ig) all from Proteintech Group, Inc. for 16 h at 4 ̊C.

Techniques: Flow Cytometry, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 1 CBX3 expression in melanoma and its correlation with disease prognosis. (A) Upregulation of CBX3 in most cancers was confirmed in the GEPIA database. (B) CBX3 expression was significantly increased in melanoma compared to its level in normal tissues (102 primary tumor samples vs. 811 normal samples, p < 0.001). (C–E) Survival curves of patient groups differentiated by CBX3 expression level from TCGA, GSE133713, and GSE19234 datasets. (F) Representative image of CBX3/HP1γ staining (high/low expression) in tumor cell nuclei (left × 40, right × 400)

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Expressing, Staining

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 2 CBX3 expression in melanoma cell lines. (A) CBX3 expression in skin cancer cell lines (n = 62). (B, C) CBX3 mRNA expression in A375, Mewo, and A2058 cell lines was explored by qPCR and western blot. (D) Overexpression of CBX3 (oe) in a transfected A2058 cell line and knockout of CBX3 (sh) in transfected A375 and Mewo cell lines in comparison with CBX3 levels in the cells transfected with the control vector (nc) were verified by western blotting. All full-length blots are presented in Supplementary file 1. (n = 3, *p < 0.05, **p < 0.01)

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Expressing, Western Blot, Over Expression, Transfection, Knock-Out, Comparison, Control, Plasmid Preparation

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 3 CBX3 promotes proliferation and migration of melanoma cells. (A) Growth rates of melanoma cells with altered CBX3 expression were determined using the CCK8 assay. (B) Tumorigenicity of melanoma cells with altered CBX3 expression was measured using the colony assay. (C, D) Migration ability of melanoma cells with altered CBX3 expression was assessed using the wound healing and transwell assays. (n = 3, *p < 0.05, **p < 0.01)

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Migration, Expressing, CCK-8 Assay, Colony Assay

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 4 CBX3 promotes tumor formation in xenograft mice. (A, B) Growth of melanoma tumors in xenograft nude mice depending on CBX3 expression levels in implanted cells. (C) Relationship between CBX3 expression and tumor volume in vivo. (D) Relationship between CBX3 expression and tumor weight in vivo. (n = 3, *p < 0.05, **p < 0.01)

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Expressing, In Vivo

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 5 Regulation of cell cycle by CBX3. (A) Differential gene expression analysis based on the CBX3 expression level. (B, C) Functional enrichment of differentially expressed genes. (D) Analysis of cell cycle in cells with different levels of CBX3 expression was performed using flow cytometry

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Gene Expression, Expressing, Functional Assay, Flow Cytometry

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 6 Regulation of cell cycle by CBX3. (A) Expression levels of hallmark cell cycle proteins were determined using western blotting. (B) In vivo expression of cell cycle hallmark proteins. All full-length blots are presented in Supplementary file 1. (C–E) The 3D protein–protein interaction models created by HDOCK and pymol. Model 1 (C), model 2 (D), and model 4 (E) were selected based on the Docking Score, Confidence Score, and Ligand rmsd. All full-length blots are presented in Supplementary file 1. (n = 3, *p < 0.05, **p < 0.01)

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Expressing, Western Blot, In Vivo

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 7 Relationships between CBX3 expression and abundances of different immune cell populations in melanoma. (A) Single-cell sequencing (sc-seq) data from dataset GSE72056. (B) CBX3 expression in sc-seq data. (C) Directions of differentiation in sc-seq data determined by the pseudotime analysis. (D) Relationship between CBX3 expression coefficient and immune cell infiltration determined by the “CIBERSORT” package. (E) CBX3 expression levels in the cancer-immunity cycle

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Expressing, Sequencing

Journal: Stem cell research & therapy

Article Title: Induction of the p21/CDK6 pathway and alteration of the immune microenvironment by the stem cell marker CBX3 in melanoma.

doi: 10.1186/s13287-025-04179-8

Figure Lengend Snippet: Fig. 8 Hypothetical mechanism of cell cycle regulation by CBX3 in melanoma. CBX3 suppressed p21 expression, disrupted the inhibitory effect of p21 on the formation of the cell cycle protein kinase complex (cyclin D1–CDK6) and promoted the G1/S phase transition. Induced by mitogenic signals, cyclin D binds to CDK6 and promotes the phosphorylation of the retinoblastoma protein (RB), which separates the transcription factor E2F from the RB1– E2F1 complex, leading to the entry of the cell into the S phase and initiation of DNA replication

Article Snippet: The primary antibodies including CBX3 (catalog number: 11650–2-AP), CDK6 (catalog number: 14052– 1-AP), p21 (catalog number: 10355–1-AP) and GAPDH (catalog number: 10494–1-AP) were offered by Proteintech (Shanghai, China).

Techniques: Expressing, Sublimation, Phospho-proteomics

Journal: Oncology reports

Article Title: [6]-Gingerol enhances the radiosensitivity of gastric cancer via G2/M phase arrest and apoptosis induction.

doi: 10.3892/or.2018.6292

Figure Lengend Snippet: Figure 4. Effects of [6]-gingerol on IR-induced cell cycle regulatory proteins and p27 mRNA expression. HGC-27 cells were treated with the vehicle, [6]-gin- gerol (300 µM), 4 Gy of IR alone, or exposed to IR (4 Gy) post [6]-gingerol (300 µM) incubation. (A) After treatment, the cells were harvested and total cell lysates were subjected to western blotting. The levels of cyclin D1, cyclin B1, cyclin A2, CDC2, CDK6 and β-actin were analyzed. Typical images of three independent experiments were presented. (B-F) Statistical analysis of the protein expression levels. (G) The relative mRNA level of p27 in the indicated treatments was assessed by qRT-PCR. *Significant difference between the indicated groups; **P<0.01, *P<0.05. #Significant difference between the IR and [6]-gingerol+IR treatment groups; ##P<0.01, #P<0.05. IR, ionizing radiation; NS, no statistical significance.

Article Snippet: Primary antibodies against cyclin B1 (cat. no. 55004-1-AP), CDK6 (cat. no. 14052-1-AP), β-actin (cat. no. HRP-60008) and tubulin (cat. no. HRP-66031) were purchased from ProteinTech Group, Inc. (Chicago, IL, USA).

Techniques: Expressing, Incubation, Western Blot, Quantitative RT-PCR