rabbit polyclonal anti tp63  (Cell Signaling Technology Inc)

Journal: Scientific reports

Article Title: Expression of p53, p63, podoplanin and Ki-67 in recurring versus non-recurring oral leukoplakia.

doi: 10.1038/s41598-021-99326-5

Figure Lengend Snippet: Figure 1. Representative images for immunohistochemical expression of Ki-67 (A), p63 (B), p53 (C) and Podoplanin (D) in leukoplakia. QuPath: Open source software for digital pathology image analysis (version 0.2.0-m4)46 was used to create the figure.

Article Snippet: Antibodies and clones Dilutions PDPN (D2-40)—M361901-2, clone D2-40, DAKO 1:150 Monoclonal mouse anti-human p53 primary antibody—M700101-2, Clone DO-7, DAKO 1:1000 Polyclonal rabbit antibody anti-human p63 primary antibody—HPA 006288, ATLAS ANTIBODIES, Sweden 1:100 Monoclonal mouse anti-human Ki-67—Clone MIB-1, DAKO 1:150 10 Vol:. (1234567890) Scientific Reports | (2021) 11:20781 | https://doi.org/10.1038/s41598-021-99326-5 were completed.

Techniques: Immunohistochemical staining, Expressing, Software

Journal: Scientific reports

Article Title: Expression of p53, p63, podoplanin and Ki-67 in recurring versus non-recurring oral leukoplakia.

doi: 10.1038/s41598-021-99326-5

Figure Lengend Snippet: Figure 2. Representative images for the immunoexpression and boxplots for percentage of positive cells in recurring versus non-recurring leukoplakia for Ki-67 (A), p63 (B), p53 (C) and podoplanin (d240) (D). Box plot graphs showed median (horizontal bars), interquartile range, and maximum and minimum values (whiskers). QuPath: Open source software for digital pathology image analysis (version 0.2.0-m4)46 and Stata Statistical Software (release 16) were used to create the figure.

Article Snippet: Antibodies and clones Dilutions PDPN (D2-40)—M361901-2, clone D2-40, DAKO 1:150 Monoclonal mouse anti-human p53 primary antibody—M700101-2, Clone DO-7, DAKO 1:1000 Polyclonal rabbit antibody anti-human p63 primary antibody—HPA 006288, ATLAS ANTIBODIES, Sweden 1:100 Monoclonal mouse anti-human Ki-67—Clone MIB-1, DAKO 1:150 10 Vol:. (1234567890) Scientific Reports | (2021) 11:20781 | https://doi.org/10.1038/s41598-021-99326-5 were completed.

Techniques: Software

Journal: Scientific reports

Article Title: Expression of p53, p63, podoplanin and Ki-67 in recurring versus non-recurring oral leukoplakia.

doi: 10.1038/s41598-021-99326-5

Figure Lengend Snippet: Figure 3. Kaplan–Meier analyses charting the cumulative probability of recurrence of leukoplakia divided by the median expression of Ki-67 (A), p63 (B), p53 (C) and podoplanin (D). Graphs show a trend that the high expression group of all four biomarkers by themselves are associated with recurrence after surgical excisions. SPSS Statistic (for Macintosh ver. 25.0 software package) was used to create the figure.

Article Snippet: Antibodies and clones Dilutions PDPN (D2-40)—M361901-2, clone D2-40, DAKO 1:150 Monoclonal mouse anti-human p53 primary antibody—M700101-2, Clone DO-7, DAKO 1:1000 Polyclonal rabbit antibody anti-human p63 primary antibody—HPA 006288, ATLAS ANTIBODIES, Sweden 1:100 Monoclonal mouse anti-human Ki-67—Clone MIB-1, DAKO 1:150 10 Vol:. (1234567890) Scientific Reports | (2021) 11:20781 | https://doi.org/10.1038/s41598-021-99326-5 were completed.

Techniques: Expressing, Software

Journal: Scientific reports

Article Title: Expression of p53, p63, podoplanin and Ki-67 in recurring versus non-recurring oral leukoplakia.

doi: 10.1038/s41598-021-99326-5

Figure Lengend Snippet: Figure 4. Kaplan–Meier analysis showing the cumulative probability of recurrence of leukoplakia. Leukoplakia were divided into high and low expression groups using the combined expression of p53 and p63 as described in the method section. The high expression group was found to have more recurrence hazards as compared to the low expression group (Log Rank, (p = 0.036). Stata Statistical Software (release 16) was used to create the figure.

Article Snippet: Antibodies and clones Dilutions PDPN (D2-40)—M361901-2, clone D2-40, DAKO 1:150 Monoclonal mouse anti-human p53 primary antibody—M700101-2, Clone DO-7, DAKO 1:1000 Polyclonal rabbit antibody anti-human p63 primary antibody—HPA 006288, ATLAS ANTIBODIES, Sweden 1:100 Monoclonal mouse anti-human Ki-67—Clone MIB-1, DAKO 1:150 10 Vol:. (1234567890) Scientific Reports | (2021) 11:20781 | https://doi.org/10.1038/s41598-021-99326-5 were completed.

Techniques: Expressing, Software

Journal: Theranostics

Article Title: Sulforaphane Inhibits the Acquisition of Tobacco Smoke-Induced Lung Cancer Stem Cell-Like Properties via the IL-6/ΔNp63α/Notch Axis

doi: 10.7150/thno.33812

Figure Lengend Snippet: ΔNp63α upregulation correlates with TS-associated lung CSC-like properties. (A) Western blot analysis of ΔNp63α, CD133, and Oct4 protein expression in lung cancer tissues. (B) Densitometry analysis (fold-change) of western blot after β-tubulin normalization (n = 12 per group). (C and D) The Pearson correlation coefficient ( r ) was used to analyze the relationship between ΔNp63α protein expression and the levels of CD133 (C) and Oct4 (D) in lung cancer specimens (n = 24). Data are expressed as the mean ± SD. Significance was assessed by unpaired two-tailed Student's t tests. ** P < 0.01 compared to the non-smoker group.

Article Snippet: The membranes were blocked with 5 % nonfat dry milk at 25 °C for 1 h on a rotary shaker and incubated overnight at 4 °C with the following primary antibodies: ΔNp63α (TA327976; OriGene Technologies, Rockville, MD, USA); CD133 (18470-1-AP), ALDH1A1 (15910-1-AP), Nanog (14295-1-AP), Oct4 (60242-1-Ig), Sox2 (11064-1-AP), ZO-1 (21773-1-AP), E-cadherin (20874-1-AP), Vimentin (10366-1-AP), N-cadherin (22018-1-AP), IL-6 (21865-1-AP), and β-tubulin (10094-1-AP; Proteintech, Rosemont, IL, USA); NICD (ab83232) and Hes1 (ab119776; Abcam, Cambridge, Massachusetts, US); β-actin (AP0060; Biogot Technology, Nanjing, China).

Techniques: Western Blot, Expressing, Two Tailed Test

Journal: Theranostics

Article Title: Sulforaphane Inhibits the Acquisition of Tobacco Smoke-Induced Lung Cancer Stem Cell-Like Properties via the IL-6/ΔNp63α/Notch Axis

doi: 10.7150/thno.33812

Figure Lengend Snippet: ΔNp63α promotes the TS-induced acquisition of CSC-like properties in HBE cells. (A) ΔNp63α expression was measured in CHBE and THBE sphere-forming cells. (B) Densitometric analyses of western blots for ΔNp63α were conducted after β-actin normalization. THBE sphere-forming cells were transfected with a control vector, ΔNp63α plasmids, control-siRNA, or ΔNp63α-siRNA for 4 days. Representative tumorsphere images were acquired (C) (original magnification, 400×) and the number of tumorspheres was quantified (D) . (E) Western blotting of ΔNp63α, CD133, ALDH1A1, and Sox2 was performed, and (F) the relative protein levels were determined in the presence or absence of ΔNp63α plasmid transfection. THBE sphere-forming cells were treated with or without ΔNp63α-siRNA for 4 days and the expression of the indicated proteins was detected by western blot analyses (G) . (H) Densitometric analyses of western blots of ΔNp63α, CD133, ALDH1A1, and Sox2 were performed after β-actin normalization. (I) Immunofluorescence staining of CD133 and ALDH1A1 in THBE sphere-forming cells for the indicated groups. Three independent experiments were performed; original magnification, 200×. Data are expressed as the mean ± SD. Significance was assessed by unpaired two-tailed Student's t tests. * P < 0.05, ** P < 0.01 compared to the vector or control-siRNA group.

Article Snippet: The membranes were blocked with 5 % nonfat dry milk at 25 °C for 1 h on a rotary shaker and incubated overnight at 4 °C with the following primary antibodies: ΔNp63α (TA327976; OriGene Technologies, Rockville, MD, USA); CD133 (18470-1-AP), ALDH1A1 (15910-1-AP), Nanog (14295-1-AP), Oct4 (60242-1-Ig), Sox2 (11064-1-AP), ZO-1 (21773-1-AP), E-cadherin (20874-1-AP), Vimentin (10366-1-AP), N-cadherin (22018-1-AP), IL-6 (21865-1-AP), and β-tubulin (10094-1-AP; Proteintech, Rosemont, IL, USA); NICD (ab83232) and Hes1 (ab119776; Abcam, Cambridge, Massachusetts, US); β-actin (AP0060; Biogot Technology, Nanjing, China).

Techniques: Expressing, Western Blot, Transfection, Control, Plasmid Preparation, Immunofluorescence, Staining, Two Tailed Test

Journal: Theranostics

Article Title: Sulforaphane Inhibits the Acquisition of Tobacco Smoke-Induced Lung Cancer Stem Cell-Like Properties via the IL-6/ΔNp63α/Notch Axis

doi: 10.7150/thno.33812

Figure Lengend Snippet: ΔNp63α transcriptionally regulates the effect of the Notch pathway on TS-induced lung CSC-like properties. (A and C) THBE sphere-forming cells were transfected with ΔNp63α plasmids, a control vector, siRNA-ΔNp63α, or control- siRNA for 4 days and the protein levels of NICD and Hes1 were determined by western blotting. (B and D) Densitometry results are shown as the fold-change compared to the vector or control siRNA after β-actin normalization. (E and F) THBE sphere-forming cells were co-transfected with wt-Notch1 promoter-luciferase, ΔNp63α (E) , or ΔNp63α-siRNA (F) , and luciferase activity was measured after incubating for 3 days. (G and H) THBE sphere-forming cells were co-transfected with mut-Notch1 promoter-luciferase, ΔNp63α (G) , or ΔNp63α siRNA (H) , and their luciferase activity was measured. Three independent experiments were performed. Data are expressed as the mean ± SD. Significance was assessed by unpaired two-tailed Student's t tests. * P < 0.05, compared to the vector group. # P < 0.05, compared to the control siRNA group.

Article Snippet: The membranes were blocked with 5 % nonfat dry milk at 25 °C for 1 h on a rotary shaker and incubated overnight at 4 °C with the following primary antibodies: ΔNp63α (TA327976; OriGene Technologies, Rockville, MD, USA); CD133 (18470-1-AP), ALDH1A1 (15910-1-AP), Nanog (14295-1-AP), Oct4 (60242-1-Ig), Sox2 (11064-1-AP), ZO-1 (21773-1-AP), E-cadherin (20874-1-AP), Vimentin (10366-1-AP), N-cadherin (22018-1-AP), IL-6 (21865-1-AP), and β-tubulin (10094-1-AP; Proteintech, Rosemont, IL, USA); NICD (ab83232) and Hes1 (ab119776; Abcam, Cambridge, Massachusetts, US); β-actin (AP0060; Biogot Technology, Nanjing, China).

Techniques: Transfection, Control, Plasmid Preparation, Western Blot, Luciferase, Activity Assay, Two Tailed Test

Journal: Theranostics

Article Title: Sulforaphane Inhibits the Acquisition of Tobacco Smoke-Induced Lung Cancer Stem Cell-Like Properties via the IL-6/ΔNp63α/Notch Axis

doi: 10.7150/thno.33812

Figure Lengend Snippet: IL-6 increases ΔNp63α in cells with TS exposure-induced lung CSC-like properties. (A) IL-6 levels in the lung cancer tissues of smokers and non-smokers were analyzed by ELISA (n = 12 per group). (B) IL-6 expression was significantly and positively correlated with ΔNp63α expression in the lung cancer specimens (Pearson correlation coefficient analysis). (C-F) THBE sphere-forming cells were exposed to various IL-6 concentrations for 7 days. The size (original magnification, 400×) (C) , and number (D) of tumorspheres were measured. The expression levels of ΔNp63α and lung CSCs markers were measured by western blotting (E) and densitometric analyses (F). Three independent experiments were performed. Data are expressed as the mean ± SD. Significance was assessed by unpaired two-tailed student's t tests or one-way ANOVA test. * P < 0.05, ** P < 0.01, compared to the control group.

Article Snippet: The membranes were blocked with 5 % nonfat dry milk at 25 °C for 1 h on a rotary shaker and incubated overnight at 4 °C with the following primary antibodies: ΔNp63α (TA327976; OriGene Technologies, Rockville, MD, USA); CD133 (18470-1-AP), ALDH1A1 (15910-1-AP), Nanog (14295-1-AP), Oct4 (60242-1-Ig), Sox2 (11064-1-AP), ZO-1 (21773-1-AP), E-cadherin (20874-1-AP), Vimentin (10366-1-AP), N-cadherin (22018-1-AP), IL-6 (21865-1-AP), and β-tubulin (10094-1-AP; Proteintech, Rosemont, IL, USA); NICD (ab83232) and Hes1 (ab119776; Abcam, Cambridge, Massachusetts, US); β-actin (AP0060; Biogot Technology, Nanjing, China).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Two Tailed Test, Control

Journal: Theranostics

Article Title: Sulforaphane Inhibits the Acquisition of Tobacco Smoke-Induced Lung Cancer Stem Cell-Like Properties via the IL-6/ΔNp63α/Notch Axis

doi: 10.7150/thno.33812

Figure Lengend Snippet: IL-6/ΔNp63α/Notch axis mediates the suppression of long-term TS exposure-induced lung CSC-like properties by SFN. (A) THBE tumorspheres were treated with different SFN concentrations (0, 5, 10, and 15 µM) for 7 days. The protein expression levels of IL-6 and ΔNp63α were detected by western blotting. (B) The levels of the indicated proteins relative to β-actin were assessed by densitometric analysis. (C) THBE sphere-forming cells were pretreated with SFN (15 μM) for 3 days, stimulated by IL-6 (20 ng/mL) for 24 h, then ΔNp63α and CD133 expression were detected by western blotting. (D) Densitometry results are shown as the fold-change compared to the control group after β-actin normalization. (E) Tumorspheres formed by THBE cells were transfected with ΔNp63α plasmids, cultured in the presence or absence of 15 µM SFN for 4 days, and the protein levels of ΔNp63α and lung CSCs markers were then measured by western blotting. (F) Densitometry results are shown as the fold-change compared to the control group after β-actin normalization. (G) Immunofluorescence staining of CD133, ALDH1A1, and Nanog in THBE sphere-forming cells for the indicated groups, original magnification, 200×. (H) The luciferase activity of the Notch1 promoter was measured in the THBE sphere-forming cells after 4 days of incubation with SFN. (I) Western blot analyses were performed, and (J) the relative protein levels of NICD and Hes1 were determined. (K) NICD and Hes1 expression in ΔNp63α transfected THBE sphere-forming cells with or without of SFN (15 µM) treatment. (L) Densitometric analyses of western blots of NICD and Hes1 were performed after β-actin normalization. (M) Immunofluorescence staining of NICD and Hes1 in THBE sphere-forming cells for the indicated groups, original magnification, 200×. Three independent experiments were performed. Data are expressed as the mean ± SD. Significance was assessed by one-way ANOVA or unpaired two-tailed Student's t tests. * P < 0.05, ** P < 0.01 compared to the control group; # P < 0.05 compared to the SFN treatment group.

Article Snippet: The membranes were blocked with 5 % nonfat dry milk at 25 °C for 1 h on a rotary shaker and incubated overnight at 4 °C with the following primary antibodies: ΔNp63α (TA327976; OriGene Technologies, Rockville, MD, USA); CD133 (18470-1-AP), ALDH1A1 (15910-1-AP), Nanog (14295-1-AP), Oct4 (60242-1-Ig), Sox2 (11064-1-AP), ZO-1 (21773-1-AP), E-cadherin (20874-1-AP), Vimentin (10366-1-AP), N-cadherin (22018-1-AP), IL-6 (21865-1-AP), and β-tubulin (10094-1-AP; Proteintech, Rosemont, IL, USA); NICD (ab83232) and Hes1 (ab119776; Abcam, Cambridge, Massachusetts, US); β-actin (AP0060; Biogot Technology, Nanjing, China).

Techniques: Expressing, Western Blot, Control, Transfection, Cell Culture, Immunofluorescence, Staining, Luciferase, Activity Assay, Incubation, Two Tailed Test

Journal: Theranostics

Article Title: Sulforaphane Inhibits the Acquisition of Tobacco Smoke-Induced Lung Cancer Stem Cell-Like Properties via the IL-6/ΔNp63α/Notch Axis

doi: 10.7150/thno.33812

Figure Lengend Snippet: SFN suppresses the IL-6/ΔNp63α/Notch axis to inhibit TS-induced lung CSC-like properties. THBE cells (1 × 10 6 ) were injected subcutaneously into the flank of nude mice. Approximately 10 days later, when palpable tumors were present, the mice were randomly administered either SFN (25 or 50 mg/kg body weight) or a vehicle control for 3 weeks, then tumor size (A) and weight (B) were measured. (C) Western blotting and (D) the relative protein levels of IL-6, ΔNp63α, NICD, Hes1, and lung CSC markers were determined. Five animal samples per group were used for densitometric analysis. (E) Schematic diagram showing the mechanism involved in the TS-mediated induction of lung CSCs. Exposure of human bronchial epithelial HBE cells to CSE induces the acquisition of lung CSC-like properties by activating ΔNp63α/Notch via IL-6 upregulation. SFN inhibits TS-triggered lung CSC-like properties via the IL-6/ΔNp63α/Notch axis. Data are expressed as the mean ± SD. Significance was assessed by one-way ANOVA or unpaired two-tailed Student's t tests. * P < 0.05, ** P < 0.01 compared to the control group.

Article Snippet: The membranes were blocked with 5 % nonfat dry milk at 25 °C for 1 h on a rotary shaker and incubated overnight at 4 °C with the following primary antibodies: ΔNp63α (TA327976; OriGene Technologies, Rockville, MD, USA); CD133 (18470-1-AP), ALDH1A1 (15910-1-AP), Nanog (14295-1-AP), Oct4 (60242-1-Ig), Sox2 (11064-1-AP), ZO-1 (21773-1-AP), E-cadherin (20874-1-AP), Vimentin (10366-1-AP), N-cadherin (22018-1-AP), IL-6 (21865-1-AP), and β-tubulin (10094-1-AP; Proteintech, Rosemont, IL, USA); NICD (ab83232) and Hes1 (ab119776; Abcam, Cambridge, Massachusetts, US); β-actin (AP0060; Biogot Technology, Nanjing, China).

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

Journal: Cell reports

Article Title: TP63-Mediated Enhancer Reprogramming Drives the Squamous Subtype of Pancreatic Ductal Adenocarcinoma

doi: 10.1016/j.celrep.2018.10.051

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-TP63 (for WB, IHC and ChIP) , Cell Signaling , Cat# 39692; RRID: N/A.

Techniques: Plasmid Preparation, Recombinant, Sample Prep, SYBR Green Assay, Reverse Transcription, Purification, Gel Extraction, Cell Viability Assay, Membrane, Microarray, Software