Journal: International Journal of Clinical and Experimental Pathology

Article Title: PinX1 is up-regulated and associated with poor patients’ survival in gliomas

doi:

Figure Lengend Snippet: Relationship between PinX1 staining and clinicopathological characteristics of the individuals in two cohorts of glioma patients

Article Snippet: The anti-PinX1 antibody (1:50 dilution, Novus Biologicals, Littleton, USA) was used for primary antibody incubation at 4°C overnight.

Techniques: Staining, Biomarker Discovery

Journal: International Journal of Clinical and Experimental Pathology

Article Title: PinX1 is up-regulated and associated with poor patients’ survival in gliomas

doi:

Figure Lengend Snippet: Immunohistochemical analysis of PinX1 expression in normal brain tissue (NB), tumor adjacent normal brain tissue (AB), benign tumor (BT), and malignant tumor (MT). (A) Low PinX1 nuclear staining in normal brain tissue; (B) Low PinX1 nuclear staining in adjacent normal brain tissue; (C) High PinX1 nuclear staining in benign tumor; (D) High PinX1 nuclear staining in malignant tumor. Magnification × 100.

Article Snippet: The anti-PinX1 antibody (1:50 dilution, Novus Biologicals, Littleton, USA) was used for primary antibody incubation at 4°C overnight.

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: International Journal of Clinical and Experimental Pathology

Article Title: PinX1 is up-regulated and associated with poor patients’ survival in gliomas

doi:

Figure Lengend Snippet: PinX1 expression was increased in the gliomas tissues when compared with the normal brain tissues (P = 0.001, Fisher’s exact test), and further increased in malignant gliomas tissues when compared with benign gliomas tissues (P = 0.090, Fisher’s exact test).

Article Snippet: The anti-PinX1 antibody (1:50 dilution, Novus Biologicals, Littleton, USA) was used for primary antibody incubation at 4°C overnight.

Techniques: Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: PinX1 is up-regulated and associated with poor patients’ survival in gliomas

doi:

Figure Lengend Snippet: Kaplan-Meier survival analyses of glioma patients. A. High PinX1 expression correlates with a poorer 5-year overall survival (P = 0.016, log-rank test). B. High PinX1 expression correlates with a poorer 5-year disease-specific survival (P=0.026, log-rank test). Abbreviation: Cum, cumulative.

Article Snippet: The anti-PinX1 antibody (1:50 dilution, Novus Biologicals, Littleton, USA) was used for primary antibody incubation at 4°C overnight.

Techniques: Expressing

Journal: International Journal of Clinical and Experimental Pathology

Article Title: PinX1 is up-regulated and associated with poor patients’ survival in gliomas

doi:

Figure Lengend Snippet: Univariate Cox proportional regression analysis on 5-year overall and disease-specific survival of 136 glioma patients

Article Snippet: The anti-PinX1 antibody (1:50 dilution, Novus Biologicals, Littleton, USA) was used for primary antibody incubation at 4°C overnight.

Techniques:

Journal: International Journal of Clinical and Experimental Pathology

Article Title: PinX1 is up-regulated and associated with poor patients’ survival in gliomas

doi:

Figure Lengend Snippet: Multivariate Cox regression analysis on 5-year overall and disease-specific survival of 136 glioma patients

Article Snippet: The anti-PinX1 antibody (1:50 dilution, Novus Biologicals, Littleton, USA) was used for primary antibody incubation at 4°C overnight.

Techniques:

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 1. PinX1 expression was observed in 26 pairs of breast cancer tissues and the normal counterparts. (A and B) Protein levels of PinX1, as assessed by immunoblot analysis with an anti-PinX1 antibody are downregulated in a majority of cancer tissues compared with the paired normal breast tissues (23/26). Separately, (B) showed significant differences (**P<0.01, two-tailed paired t-test) and (A) presented 12 pairs of the 26 pairs. GAPDH was a loading control; T, tumor; N, non-tumor. (C and D) Quantitative PCR analysis of PinX1 mRNA expression in 26 paired samples. Data are shown relative to those of the internal control gene Gapdh. (D) Significant differences (**P<0.01, two-tailed paired t-test) of PinX1 expression, (C) representing 15 pairs of the total.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Expressing, Western Blot, Two Tailed Test, Control, Real-time Polymerase Chain Reaction

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 2. Immunohistochemical staining of PinX1 was analysed in breast cancer tissues and the normal breast tissues. (A) A cancer adjacent normal breast tissue (case 1) with high expression of PinX1, where almost all tumor cells were positively stained (magnification, x100). (B) A fibroadenoma tissue (case 3) with >65% of tumor cells positive staining of PinX1 (magnification, x100). (C) An invasive ductal carcinoma (case 43) exhibited low expression of PinX1, where <45% of tumor cells were positive staining of PinX1 (magnification, x100). (G) Low expression of PinX1 was obtained in an invasive ductal carcinoma (case 29), where <10% of tumor cells were positive staining of PinX1 (magnification, x100). Respectively, (D-H) revealed the higher magnification (magnifica- tion, x400) of the specific areas boxed in (A, B, C and G).

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 3. The ROC curve was used to obtain the optimal cut-off value for positive and negative PinX1 expression. The plots of sensitivity and specificity for each clinicopathological parameter was as follows: (A) age at surgery; (B) histology grade; (C) clinical stage; (D) pT status; (E) pN status; (F) ER status; (G) PR status; (H) Her2 status; (I) p53 status.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Expressing

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 4. PinX1 inhibits proliferation in a human basal-like breast cancer cell line. (A) Immunoblot analysis of PinX1 expression levels in the PinX1- overexpressed group (PinX1-pEGFP-N1), wild-type MDA-MB-231 (WT) and negative control (pEGFP-N1). GAPDH is a loading control. (B) Immunoblot analysis of PinX1 expression levels in the PinX1-knockout group (PinX1-/-) and the wild-type control (MDA-MB-231). (C and D) The Cell Counting kit-8 (CCK-8) assay was applied to detect the proliferation rates at specific periods (0, 24, 48 and 72 h). Overexpression of PinX1 inhibits the proliferation of MDA-MB-231 while PinX1 knockout promotes its proliferation. **P<0.01.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Western Blot, Expressing, Negative Control, Control, Knock-Out, Cell Counting, CCK-8 Assay, Over Expression

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 5. PinX1 suppresses the migration ability of MDA-MB-231 cells in vitro. (A) The migration ability of the PinX1-overexpressed and the control groups were tested by wound healing assay in a 36-h recovery time (top, magnification, x20). Bottom, statistical analysis of the migration rates, presented relative to the initial wound area. (B) The migration ability of the PinX1 knockout and the wild-type groups was tested by wound healing assay in a 36-h recovery time (top, magnifications, x10). Bottom, statistical analysis of the migration rates, presented relative to the initial wound area. (C and D) Transwell assay was also used to measure the migration ability of the different experimental groups (left, magnifications, x10). Right, statistical analyses of the migrated cell number per field. At least three independent experiments were performed (*p<0.05, two-tailed unpaired t-test) and the data expressed are mean ± SEM.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Migration, In Vitro, Control, Wound Healing Assay, Knock-Out, Transwell Assay, Two Tailed Test

Journal: Oncology reports

Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer.

doi: 10.3892/or.2017.5696

Figure Lengend Snippet: Figure 6. Overexpression of PinX1 induces apoptosis of MDA-MB-231 cells in vitro. (A and B) Western blot analysis was used to measure the expression levels of cleaved caspase-3 in different experimental groups. GAPDH was a loading control. (C) Flow cytometric analysis (left) of the apoptotic rate of the PinX1- overexpressed group with its negative control and the PinX1 knockout group with its counterpart. Quantitative analyses of the apoptotic rate on the right. At least three independent experiments were preformed (*p<0.05, two-tailed unpaired t-test) and the data expressed are mean ± SEM.

Article Snippet: The blots were then blocked by 5% non-fat dry milk dissolved in Trisbuffered saline with Tween-20 (TBST) at room temperature for 1 h. Thereafter, the membranes were, respectively, incubated with primary antibodies diluted by TBST for PinX1 (1:1,000, goat anti-human polyclonal; Santa Cruz Biotechnology, Santa Cruz, CA, USA) and GAPDH (1:10,000, anti-human, conjugated with HRP; Proteintech Group, Inc., Rosemont, IL, USA) at 4 ̊C overnignt.

Techniques: Over Expression, In Vitro, Western Blot, Expressing, Control, Negative Control, Knock-Out, Two Tailed Test

Journal: Cancer cell international

Article Title: Reduced expression of PinX1 correlates to progressive features in patients with prostate cancer.

doi: 10.1186/1475-2867-14-46

Figure Lengend Snippet: Figure 1 qRT-PCR and Western blot analysis of PinX1 expression in paired PCa and adjacent normal prostate tissues. (A) Fold changes (2-△△Ct values) by qRT-PCR showed a reduced expression of PinX1 mRNA in the majority of PCa cases (14/16),when compared with paired normal prostate tissues. Expression levels were normalized for GAPDH. (B) Significant differences of PinX1 mRNA expression between the PCa and adjacent normal prostate tissues (P < 0.0001). (C) Western blotting indicated down-regulation of PinX1 protein in PCa tissues (15/16) in comparison with the adjacent normal prostate tissues. β-actin was used as internal control. T, PCa; N, normal. (D) Significant differences of PinX1 protein expression between the PCa and adjacent normal prostate tissues (P < 0.0001).

Article Snippet: Goat polyclonal anti-PinX1 antibody (1:500, Santa Cruz Biotechnology, Santa Cruz, USA) and HRP conjugated rabbit anti-goat IgG (1:5000, Multisciences, Hangzhou, China) were used to detect the PinX1 protein.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Comparison, Control

Journal: Cancer cell international

Article Title: Reduced expression of PinX1 correlates to progressive features in patients with prostate cancer.

doi: 10.1186/1475-2867-14-46

Figure Lengend Snippet: Figure 2 The expression of PinX1 protein in normal prostate and PCa tissues by IHC on the TMA. (A) Positive expression of PinX1 in normal prostate tissue, in which about 100% of the prostate epithelial cells demonstrated a strong nuclear staining of PinX1, while a lesser staining in the cytoplasm could also be observed (100x). (B) Positive expression of PinX1 in PCa (case 21), in which more than 90% of tumor cells showed positive staining (100x). (C) Negative expression of PinX1 in PCa (case 11), with less than 40% positive tumor cells (100x). (D) Negative expression of PinX1 in PCa (case 27), with less than 35% positive tumor cells (100x). (E) Negative expression of PinX1 in PCa (case 25), with less than 15% positive tumor cells (100x). (F), (G), (H), (I) and (J) demonstrate the higher magnification (400x) from the area of black square in (A), (B), (C), (D) and (E), respectively.

Article Snippet: Goat polyclonal anti-PinX1 antibody (1:500, Santa Cruz Biotechnology, Santa Cruz, USA) and HRP conjugated rabbit anti-goat IgG (1:5000, Multisciences, Hangzhou, China) were used to detect the PinX1 protein.

Techniques: Expressing, Staining

Journal: Cancer cell international

Article Title: Reduced expression of PinX1 correlates to progressive features in patients with prostate cancer.

doi: 10.1186/1475-2867-14-46

Figure Lengend Snippet: Figure 3 Selection of the optimum cut-off score for positive expression of PinX1 by receiver operating characteristic (ROC) analysis. Various ROC curves were plotted by sensitivity and specificity for each outcome: age (A), histological grade (B), clinical stage (C), Gleason pattern (D), Gleason grade (E), pT status (F), pN status (G), pM status (H).

Article Snippet: Goat polyclonal anti-PinX1 antibody (1:500, Santa Cruz Biotechnology, Santa Cruz, USA) and HRP conjugated rabbit anti-goat IgG (1:5000, Multisciences, Hangzhou, China) were used to detect the PinX1 protein.

Techniques: Selection, Expressing

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 1. Immunostaining of PinX1 in CRC tissues. (A) Intensity of staining in CRC tissues. Top panel, x100 magnification; bottom panel, x200 magnification; a and e, negative; b and f, weak; c and g, moderate; d and h, strong. (B) The distribution of different staining intensities of PinX1 in CRC tissues compared with adjacent non‑cancerous control tissues (P<0.001, paired Wilcoxon signed‑rank test). (C) Low PinX1 expression is associated with lymph node metastasis (*P=0.021, χ2 test). (D) Low PinX1 expression is associated with distant metastasis (*P=0.030, χ2 test). (E) Low PinX1 expression is associated with advanced Tumor‑Node‑Metastasis stage (*P=0.014, χ2 test). PinX1, Pin2/TRF1‑binding protein X1; CRC, colorectal cancer; N‑C, adjacent non‑cancerous tissues‑CRC tissues; N, node; M, metastasis.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Immunostaining, Staining, Control, Expressing

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 2. Expression of PinX1 is associated with overall and disease‑free survival in patients with CRC. (A) Low PinX1 expression is associated with a poorer overall cumulative survival in patients with CRC (P=0.001, log‑rank test). (B) Low PinX1 expression is associated with a poorer disease‑free cumulative survival in patients with CRC (P=0.017, log‑rank test). Cum, cumulative; PinX1, Pin2/TRF1‑binding protein X1.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Expressing

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 3. PinX1 inhibits the migration and invasion of colorectal cancer cells. (A) The relative protein expression level of PinX1 in PinX1‑knockdown (shPinX1) and control groups (shCtrl) in HCT116 and SW480 cells was detected by western blotting. (B) Densitometric analysis of the relative protein expression level of PinX1 in PinX1‑knockdown (shPinX1) and control groups (shCtrl) in HCT116 and SW480 cells. (C and D) PinX1‑knockdown inhibited the migration ability of HCT116 and SW480 cells. (E and F) PinX1‑knockdown inhibited the invasion ability of HCT116 and SW480 cells. (G and H) PinX1‑knockdown inhibited the proliferation ability of HCT116 and SW480 cells. All experiments were performed in triplicate. Data are presented as the mean ± standard deviation. ***P<0.001; **P<0.01. PinX1, Pin2/TRF1‑binding protein X1; sh, short hairpin RNA; Ctrl, control.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Migration, Expressing, Control, Western Blot, Standard Deviation, shRNA

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 4. PinX1 expression inhibits the expression and activity of MMP2, and inhibits expression of p65 and p‑p65. (A) Western blot analysis of the relative protein levels of PinX1, MMP2, MMP9, TIMP‑1 and TIMP‑2 in HCT116 and SW480. (B) Gelatin zymography analysis of MMP2 in HCT116 and SW480. (C) Western blot analysis of the relative protein levels of PinX1, MMP2, p65 and p‑p65 in HCT116 and SW480. All experiments were performed in triplicate through comparing knockdown of the PinX1 group (shPinX1) with that of the control group (shCtrl). Histograms represent the mean ± standard deviation. ***P<0.001. PinX1, Pin2/TRF1‑binding protein X1; MMP2, matrix metalloproteinase 2; p‑, phosphorylated; TIMP, TMP metallopeptidase inhibitor 1; sh, short hairpin RNA; Ctrl, control.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Expressing, Activity Assay, Western Blot, Zymography, Knockdown, Control, Standard Deviation, shRNA

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 5. PinX1 suppresses MMP2 expression via the nuclear factor‑κB pathway. Western blot analysis of the relative protein expression levels of PinX1, MMP2, p65 and p‑p65 in shCtrl and shPinX1 groups and those co‑treated with p65 siRNA in (A) HCT116 and (B) SW480 cells. The p65‑specific siRNA efficiently prevented the upregulation of MMP2 expression induced by the knockdown of PinX1. (C and D) The improved migration and invasion ability resulting from the knockdown of PinX1 was inhibited by p65‑siRNA in HCT116 and SW480 cells. All experiments were performed in triplicate. Histograms represent the mean ± standard deviation. **P<0.01; ***P<0.001. PinX1, Pin2/TRF1‑binding protein X1; MMP2, matrix metalloproteinase 2; p‑, phosphorylated; si, small interfering; sh, short hairpin RNA; Ctrl, control.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: Expressing, Western Blot, Knockdown, Migration, Standard Deviation, shRNA, Control

Journal: Oncology reports

Article Title: Pin2/TRF1‑binding protein X1 inhibits colorectal cancer cell migration and invasion in vitro and metastasis in vivo via the nuclear factor‑κB signaling pathway.

doi: 10.3892/or.2018.6570

Figure Lengend Snippet: Figure 6. PinX1 negatively regulates colorectal cancer cell metastasis in vivo. (A) Right panel, representative image of liver with metastatic nodules; middle panel, H&E staining sections of liver with metastatic nodules; left panel, representative image of H&E stained lung sections. (B) Representative image of 10% formalin‑fixed liver with metastatic nodules. (C) The number of liver metastatic nodules was counted under a dissecting microscope. Compared with the shCtrl group, a statistically significant increase in the number of the liver metastases was observed in the shPinX1 group (***P=0.002). (D) Immunohistochemical staining of MMP2 and p65 in metastatic liver nodules. MMP2 and p65 expression was increased in the shPinX1 group, compared with the shCtrl group. sh, short hairpin; Ctrl, control; PinX1, Pin2/TRF1‑binding protein X1; MMP2, matrix metalloproteinase 2.

Article Snippet: The slides were subsequently incubated with a polyclonal rabbit anti-PinX1 antibody (1:50; cat. no. NBP2-32265; Novus Biologicals, LLC, Littleton, CO, USA) at 4 ̊C overnight, and known immunostaining-positive/negative slides served as positive and negative controls.

Techniques: In Vivo, Staining, Microscopy, Immunohistochemical staining, Expressing, Control

Journal: iScience

Article Title: Non-random spatial organization of telomeres varies during the cell cycle and requires LAP2 and BAF

doi: 10.1016/j.isci.2024.109343

Figure Lengend Snippet:

Article Snippet: PINX1 , Bethyl , A304-389A.

Techniques: Plasmid Preparation, Recombinant, Saline, Generated, Software