Journal: Oncology Research

Article Title: Dysregulated Cell Signaling Pathways in Prostate Tumoral Plasticity—Checkpoints

doi: 10.32604/or.2026.072421

Figure Lengend Snippet: Cell cycle signaling pathway by propidium iodide (PI) stain. ( A ) Benign prostate hyperplasia (BPH) tissue samples, low S-phase category (<7%), G2/M phase arrest: G0/G1: S1 = 0.00%; S2 = 0.23%; S4 = 0.45%; S15 = 0.06%; S: S1 = 2.92%; S2 = 1.85%; S4 = 5.73%; S15 = 7.22%; G2/M: S1 = 95.83%; S2 = 96.98%; S4 = 90.82%; S15 = 91.93%. ( B ) Adenocarcinoma (AC) tissue samples, high S-phase category (>12%), G0/G1 or G2/M phases arrests, apoptosis-necrosis continuum phenotype: G0/G1: S5 = 0.37%; S6 = 30.10%; S10 = 0.21%; S22 = 47.80%; S: S5 = 13.79%; S6 = 33.02%; S10 = 15.12%; S22 = 31.04%; G2/M: S5 = 85.71%; S6 = 36.07%; S10 = 84.12%; S22 = 19.60%. ( C ) Carcinoma (CA) tissue samples, uncontrolled S-phase proliferation: G0/G1: S13 = 1.92%; S14: 0.21%; S16 = 52.51%; S31 = 4.34%; S: S13 = 73.71%; S14: 77.28%; S16 = 41.54%; S31 = 85.09%; G2/M: S13 = 22.94%; S14: 22.18%; S16 = 5.72%; S31 = 9.81%. Legend: C1DNA-negative control represented by healthy tissue sample; C2DNA-positive control represented by adenocarcinoma or carcinoma tissue samples. Negative and positive controls were extrapolated to graphs to interpret the experimental samples.

Article Snippet: Homogenized prostate tissue samples (200 μL) were fixed in absolute ethanol, in darkness, for 30 min. 10 μL of propidium iodide (PI, 00-6990, Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) were added to tubes with prostate samples (200 μL of cells at 10 6 cells/mL) and kept in the darkness at room temperature, for 30 min. 1 mL of flow cytometry staining buffer (FCB, 00-4222-26, eBioscienceTM, Invitrogen, Life Technologies Carlsbad, CA, USA) was added to the tubes.

Techniques: Staining, Negative Control, Positive Control

Journal: Nanoscale Advances

Article Title: A novel fullerene-lysine derivative with noticeable ROS scavenging capabilities for improving type 2 diabetes mellitus

doi: 10.1039/d4na01081g

Figure Lengend Snippet: Cell viability assays in vitro . (a) The apoptosis of MIN6 cells, including those treated with C 60 -Lys. (b) The percentages of apoptotic cells from a. (c) The cell cycle distributions of MIN6 cells, including after C 60 -Lys treatment. (d) The percentages of cells in each stage from c. ns means p > 0.05, * means p < 0.05, and *** means p < 0.001 compared with the control group. Data were analyzed using t -tests and ANOVA.

Article Snippet: Cell apoptosis was assessed using an Annexin V-FITC/PI apoptosis assay kit (Vazyme, China) and cell cycle was assessed using a cell cycle assay kit (Beyotime, China) according to the instructions.

Techniques: In Vitro, Control

Journal: Cell Death Discovery

Article Title: Single-cell trajectory analysis reveals a CD9 positive state to contribute to exit from stem cell-like and embryonic diapause states and transit to drug-resistant states

doi: 10.1038/s41420-023-01586-9

Figure Lengend Snippet: A Likelihood of cells remaining in the SCLED cluster based on CD9 expression. B CD9 levels of individual cells are mapped on the graph in Fig. . C Scatter plot shows Pearson correlation of average CD9 expression and cluster 9 signature in each dataset in Fig. . D Violin plot shows indicated motif activity of CD9 high cluster and CD9 low cluster in K562 and MDAMB468 cells. E Kaplan–Meier curve of relapse-free survival in patients with basal-like breast cancer ( n = 65 or n = 165) based on low or high CD9 protein split on median level. Significance is determined by log-rank test. F Changes of in vitro stemness score of SUM149 transfected with shCON or shCD9-1 lentivirus treated with JQ1 (100 nM) for 96 h, 8 days and 10 days. In vitro stemness markers included SETD6, NANOG, POU5F1, MALAT1, and NEAT1 were assessed by RT-PCR and in vitro stemness score calculated as un weighted sum of relative fold change of each gene compared to shCON cells treated with DMSO and shCD9 cells treated with DMSO, respectively. Data represent SEM of three replicates. G Changes of in vitro stemness score of SUM149 treated with JQ1 (100 nM), anti-CD9 and combination for 96 h, 8 days and 10 days. H Cell viability assessed by prestoblue of SUM149 transfected with shCON, shCD9-1, shCD9-2, and shCD9-3 lentivirus and treated with indicated doses of JQ1 for 96 h. I Scatter plot of correlation of cell growth assessed by prestoblue and CD9 expression (left). Scatter plot of cell sensitivity to JQ1 and CD9 expression (right).

Article Snippet: Antibodies used for cytometry were CD9 (C-4, Santa Cruz Biotechnology, #J0819), along with Calcein violet AM (Invitrogen, #65-0854-39), Propidium Iodide (PI) (Invitrogen, 00-6990-50).

Techniques: Expressing, Activity Assay, In Vitro, Transfection, Reverse Transcription Polymerase Chain Reaction

Journal: Cell Death Discovery

Article Title: Single-cell trajectory analysis reveals a CD9 positive state to contribute to exit from stem cell-like and embryonic diapause states and transit to drug-resistant states

doi: 10.1038/s41420-023-01586-9

Figure Lengend Snippet: A Representative light microscopy images of 3D cultured cell lines treated with indicated drug for 96 h. (JQ1 = 100 nM, MG132 = 400 nM, mTORi = 200 nM, HSP70i = 10 μM, HSP90i = 8 nM, cDDP=300 nM, CHK1i = 1 nM, WEE1i = 60 nM, PARPi=600 nM, Tram=500 nM, CDK4/6i = 1 μM). Data are representative of three replicates. Scale bars: 200 μm. B Bar charts of colony numbers with indicated diameter with indicated treatments in A . C Scatter plots of Pearson correlation of CD9 expression and indicated drug sensitivity in cancer cell lines from CTRPv2. Each dot presents a cell line.

Article Snippet: Antibodies used for cytometry were CD9 (C-4, Santa Cruz Biotechnology, #J0819), along with Calcein violet AM (Invitrogen, #65-0854-39), Propidium Iodide (PI) (Invitrogen, 00-6990-50).

Techniques: Light Microscopy, Cell Culture, Expressing

Journal: Cell Death Discovery

Article Title: Single-cell trajectory analysis reveals a CD9 positive state to contribute to exit from stem cell-like and embryonic diapause states and transit to drug-resistant states

doi: 10.1038/s41420-023-01586-9

Figure Lengend Snippet: A Tumor growth curves of indicated groups. SUM149 with or without stable CD9 knockdown were injected into mice. Five mice were treated with vehicle or JQ1 (25 mg/kg intraperitoneal daily) for 30 days. Tumor volume was measured with calipers. B Tumor growth curves of indicated groups. Five mice were treated with vehicle and IgG (1 mg/kg, every second day), JQ1 (10 mg/kg intraperitoneal daily), anti-CD9 antibody (1 mg/kg, every second day) or JQ1 combined with anti-CD9 antibody for 30 days. Tumor volume was measured with calipers. C scRNA data from tumors collected at 30 days was assessed and visualized using densMAP as described in Fig. . scPred trained on data from Fig. was used to provide cell cluster identity. Frequency of each treatment is shown by pie chart mapped on each cluster. D Cell distribute of each indicated treatment in densMAP. E Single cells are colored by normalized latent time, asymmetric division score, ABC score, ALDH score, embryonic diapause score, ITH score and cluster9 signature score (c9) and resistant cluster signature score (cR) mapped onto ( C ). Scale is at the left bottom of the latent time panel. F Each cell in the in vivo analysis was assigned to a SUM149 cluster using scPred trained on data from Fig. and frequency determined. G Hazard Ratio predicted by Kaplan–Meier curve of relapse-free survival in patients with basal-like breast cancer ( n = 442) based on DEGs with log 2 FC ≥ 5 and adjusted p -values < 1e−9 for clusters in ( C ) in SUM149. Significance was determined by log-rank test. * p < 0.05.

Article Snippet: Antibodies used for cytometry were CD9 (C-4, Santa Cruz Biotechnology, #J0819), along with Calcein violet AM (Invitrogen, #65-0854-39), Propidium Iodide (PI) (Invitrogen, 00-6990-50).

Techniques: Knockdown, Injection, In Vivo

Journal: Cell Death Discovery

Article Title: Single-cell trajectory analysis reveals a CD9 positive state to contribute to exit from stem cell-like and embryonic diapause states and transit to drug-resistant states

doi: 10.1038/s41420-023-01586-9

Figure Lengend Snippet: A GSVA enrichment scores of KEGG pathways in scRNA Seq of xenograft tumors from Fig. . B GSVA enrichment scores of top 50 DEGs for clusters 4.1, 4.2, 9 and 1 in SUM149. C GSVA enrichment scores of top 50 DEGs for clusters 4, 9 and 1 in SUM159. D Representative immunofluorescence images of indicated protein expression in indicated SUM149 cell lines (JQ1 = 100 nM). Data are representative of three replicates. E Representative immunofluorescence images of indicated protein expression in SUM149 cell lines treated with JQ1 (100 nM), anti-CD9 and combination. Data are representative of three replicates. F Representative western blot of data from one of three independent experiments of indicated protein in SUM149 cell line treated with indicated treatments (JQ1 = 100 nM, MG132 = 200 nM) for 96 h. G Whole-proteome thermal stability curve of SUM149 cell line treated with indicated treatments for 96 h. Data represent SEM of three replicates. H Representative western blot of indicated protein of indicated cell lines (JQ1 = 100 nM, anti-CD9 = 1 μg/μl). Data are representative of three experiments. I Representative light microscope images of 3D cultured SUM149 cell line treated with indicated drugs (JQ1 = 100 nM, MG132 = 400 nM, HSP70i = 10uM, HSP90i = 8 nM) for 96 h (left). Bar chart of colony numbers with indicated diameter with indicated treatments (right). Data represent SEM of three replicates. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Antibodies used for cytometry were CD9 (C-4, Santa Cruz Biotechnology, #J0819), along with Calcein violet AM (Invitrogen, #65-0854-39), Propidium Iodide (PI) (Invitrogen, 00-6990-50).

Techniques: Immunofluorescence, Expressing, Western Blot, Light Microscopy, Cell Culture