Journal: Frontiers in Physiology

Article Title: Adenosine stimulates the basolateral 50 pS K + channel in renal proximal tubule via adenosine-A1 receptor

doi: 10.3389/fphys.2023.1242975

Figure Lengend Snippet: Inhibition of adenosine-A2 receptor fails to abolish the effect of adenosine on the basolateral 50 pS K + channel of the PT. (A) A single channel recording (in a cell-attached patch) shows the time course of adenosine effect (1 μm) on the 50 pS K + channel in the presence of 10 μm CP-66713, an adenosine A 2 receptor antagonist. The experiments were performed in the PT pretreated with CP-66713 (which was continuously present in the bath). Two parts of the trace indicated by numbers are extended to show the fast time resolution. Holding potential is at 0 mV “C” indicates that the channel close level. (B) A Bar graph summarizes the effect of adenosine on the 50 pS K + channel in the presence of CP-66713.

Article Snippet: Collagenase II, DPCPX, CP-66713, AACOCF3, Calphostin C, U73122, H-8 (N—[2- (methylamino) ethyl] -5- isoquinoline sulfonamide-2HC1), and the chemical reagent used for preparation of above solution were all purchased from Sigma.

Techniques: Inhibition

Journal: International Journal of Biological Sciences

Article Title: Ailanthus Altissima-derived Ailanthone enhances Gastric Cancer Cell Apoptosis by Inducing the Repression of Base Excision Repair by Downregulating p23 Expression

doi: 10.7150/ijbs.60674

Figure Lengend Snippet: AIL inhibits DNA damage repair of GC cells. (A) Transcriptome sequencing and pathway enrichment were performed on PDX tumor tissues from DMSO- and AIL-treated mice. (B) Differential gene expression analysis of the transcriptome sequencing. (C) AGS, SNU719 and SGC7901 cells were treated with AIL (0.5 µM) for 0, 12, and 24 hours, the cells were then lysed, and the expression levels of P23 and XRCC1 were detected by western blotting. (D) Western blotting was performed to analyze the expression of HSP90, XRCC1, AKT and P23 in GC cell lines (AGS, SNU719 and SGC7901) after treatment with the specified concentrations of AIL for 24 hours. (E) AGS, SNU719 and SGC7901 cells were treated with 17-AAG (HSP90 inhibitor, 5 µM), CEL (P23 inhibitor, 2 µM) and AIL (1 µM) for 24 hours, and the expression of XRCC1 was detected by western blotting. (F) Representative images of P23, HSP90, and XRCC1 in PDX tumor tissue as detected via immunohistochemistry (100X). Scale bars, 200 µm. (G) Data were derived from experiments conducted in triplicate in (F). The PDX tumor tissues were compared with the control group, *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Antibodies used were: XRCC1 (1: 100, Abcam, ab44830), P23 (1: 200, Proteintech, 10824-1-AP), HSP90 (1: 400, Proteintech, 60318-1-Ig), Lamin B1 (1: 20000, Proteintech, 66095-1-Ig), GAPDH (1: 20000, Proteintech, 60004-1-Ig).

Techniques: Sequencing, Expressing, Western Blot, Immunohistochemistry, Derivative Assay

Journal: International Journal of Biological Sciences

Article Title: Ailanthus Altissima-derived Ailanthone enhances Gastric Cancer Cell Apoptosis by Inducing the Repression of Base Excision Repair by Downregulating p23 Expression

doi: 10.7150/ijbs.60674

Figure Lengend Snippet: AIL regulates XRCC1 through P23 to inhibit DNA damage repair. (A) Western blotting was performed to detect the expression of P23, HSP90 and XRCC1 in AGS, SNU719 and SGC7901 cells after treatment with different concentrations of CEL (P23 inhibitor) for 24 hours. (B) GC1, GC2, and GC3 organoids were flow-sorted to separate P23(-) and P23(+) organoid strains. (C) qPCR was performed to detect P23 mRNA levels in P23(-) and P23(+) organoid strains. (D) qPCR analysis of XRCC1 expression in P23(-) and P23(+) organoid strains. (E) Western blot analysis of the expression of HSP90 and XRCC1 in AGS, SNU719 and SGC7901 cells treated with the specified concentrations of 17-AAG (HSP90 inhibitor) for 24 hours. (F) Representative images of GC organoids treated with DMSO (control group), 17-AAG (10 µM), CEL (2 µM), and AIL (1 µM) for 14 days. The number of cell clusters (G) and size of clusters (H) were counted. Data were derived from experiments conducted in triplicate. The treated organoids were compared with the control group, *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Antibodies used were: XRCC1 (1: 100, Abcam, ab44830), P23 (1: 200, Proteintech, 10824-1-AP), HSP90 (1: 400, Proteintech, 60318-1-Ig), Lamin B1 (1: 20000, Proteintech, 66095-1-Ig), GAPDH (1: 20000, Proteintech, 60004-1-Ig).

Techniques: Western Blot, Expressing, Derivative Assay

Journal: International Journal of Biological Sciences

Article Title: Ailanthus Altissima-derived Ailanthone enhances Gastric Cancer Cell Apoptosis by Inducing the Repression of Base Excision Repair by Downregulating p23 Expression

doi: 10.7150/ijbs.60674

Figure Lengend Snippet: AIL inhibits the binding of P23 and XRCC1 to HSP90 in GC cells. (A) The colocalization of HSP90 with P23 and XRCC1 was demonstrated with immunofluorescence in GC organoids treated with AIL (1 µM) for 24 hours. Scale bars, 100 µm. (B) Data were derived from experiments conducted in triplicate in (A). (C) Pull-down of selected proteins with HSP90 in the nucleus and cytoplasm of GC cells treated with AIL for 24 hours, and then the IP fractions were immunoblotted with HSP90, XRCC1, P23, Lamin B1 and GAPDH. (D) Graphical illustration of the functions of AIL in inhibiting tumor growth. The treated cells were compared with the control group, *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Antibodies used were: XRCC1 (1: 100, Abcam, ab44830), P23 (1: 200, Proteintech, 10824-1-AP), HSP90 (1: 400, Proteintech, 60318-1-Ig), Lamin B1 (1: 20000, Proteintech, 66095-1-Ig), GAPDH (1: 20000, Proteintech, 60004-1-Ig).

Techniques: Binding Assay, Immunofluorescence, Derivative Assay

Journal: International Journal of Biological Sciences

Article Title: Ailanthus Altissima-derived Ailanthone enhances Gastric Cancer Cell Apoptosis by Inducing the Repression of Base Excision Repair by Downregulating p23 Expression

doi: 10.7150/ijbs.60674

Figure Lengend Snippet: Highly expressed P23 is associated with GC recurrence. (A) Representative images of high and low expression levels of P23 detected via IHC (100X) in recurrent GC tissues. Scale bars, 200 µm. (B) Recurrence time curves of 93 patients with GC recurrence, including 42 patients with high P23 expression and 51 patients with low P23 expression. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Antibodies used were: XRCC1 (1: 100, Abcam, ab44830), P23 (1: 200, Proteintech, 10824-1-AP), HSP90 (1: 400, Proteintech, 60318-1-Ig), Lamin B1 (1: 20000, Proteintech, 66095-1-Ig), GAPDH (1: 20000, Proteintech, 60004-1-Ig).

Techniques: Expressing

Journal: BMC Cancer

Article Title: Identification of proteins involved in neural progenitor cell targeting of gliomas

doi: 10.1186/1471-2407-9-206

Figure Lengend Snippet: Expression profiles of selected genes and proteins from microarray analysis, RT-PCR, immunocytochemistry and western blot analysis.

Article Snippet: Fluorescent immunohistochemical staining was performed by rinsing cell slides in KPBS, blocking in appropriate serum 1 h followed by primary antibody incubation; polyclonal rb anti-TPT1 (10824-1-AP Protein Tech Group, Chicago, IL), blocking goat anti-Follistatin (AF669 R&D systems, Germany), biotin conjugated hamster anti-CD81 monoclonal (TAPA1, 559518 BD Pharmingen), rb anti-AXL (4977 Cell Signaling Tec.) overnight in 5% NDS, 0.25% Triton X-100.

Techniques: Expressing, Microarray, Immunocytochemistry, Western Blot

Journal: BMC Cancer

Article Title: Identification of proteins involved in neural progenitor cell targeting of gliomas

doi: 10.1186/1471-2407-9-206

Figure Lengend Snippet: A decrease in tumor proliferation after the addition of antibodies against the genes Gas6, AXL, CXCL1, CD81 and TPT1 was observed . Co-cultures of tumor N29 (A) and N32 (B) together with irradiated NPC (HiB5/ST14A), were treated by adding antibodies against Gas6, AXL, TPT1, CXCL1 and CD81. Cells were seeded together and allowed to attach before addition of the antibodies. The plates were pulsed with 3 H-thymidine after 3 days in culture and cell division was measured as the amount of incorporated 3 H-thymidine. The result is expressed as fold change, where 1 represents the inhibition by NPC alone on the tumor (dashed line). Values below 1 show a further decrease in tumor proliferation due to the addition of antibodies. White bars show HiB5 and black bars show ST14A cultures. Mean ± stdev: Statistical analysis was made using t-test and Mann Whitney Wilcoxon rank sum test * P < 0.05

Article Snippet: Fluorescent immunohistochemical staining was performed by rinsing cell slides in KPBS, blocking in appropriate serum 1 h followed by primary antibody incubation; polyclonal rb anti-TPT1 (10824-1-AP Protein Tech Group, Chicago, IL), blocking goat anti-Follistatin (AF669 R&D systems, Germany), biotin conjugated hamster anti-CD81 monoclonal (TAPA1, 559518 BD Pharmingen), rb anti-AXL (4977 Cell Signaling Tec.) overnight in 5% NDS, 0.25% Triton X-100.

Techniques: Irradiation, Inhibition, MANN-WHITNEY