Journal: PloS one

Article Title: Reduced selenium-binding protein 1 in breast cancer correlates with poor survival and resistance to the anti-proliferative effects of selenium.

doi: 10.1371/journal.pone.0063702

Figure Lengend Snippet: Figure 1. The Expression of SELENBP1 in Normal and Tumor Breast Tissues. Breast cancer tissue arrays were stained by immunohistochemistry using anti-human SELENBP1 antibody at 1:100 dilution. Positive stained cells are shown in dark brown color. (A) Strong positive staining of SELENBP1 in normal breast tissue under low power view (200X). (B–C) Weak positive to negative staining of SELENBP1 in breast cancer tissues under high power view (400X). (D) The Allred scoring distributions of SELENBP1 expression in normal and tumor tissue groups. Inside lines represent means and standard deviations. *p,0.05. (E) Statistical results for the difference between normal and tumor tissues as analyzed by Kruskal-Wallis test. doi:10.1371/journal.pone.0063702.g001

Article Snippet: Breast Cancer Tissue Array Slides Three sets of breast cancer tissue arrays (including normal, primary tumor, and metastases) were obtained from Imgenex or Biomax Inc.

Techniques: Expressing, Staining, Immunohistochemistry, Negative Staining

Journal: PloS one

Article Title: Reduced selenium-binding protein 1 in breast cancer correlates with poor survival and resistance to the anti-proliferative effects of selenium.

doi: 10.1371/journal.pone.0063702

Figure Lengend Snippet: Figure 2. SELENBP1 Expression is Progressively Reduced in Advancing Clinical Stages in Breast Cancer Tissues. (A) The scoring distributions of SELENBP1 expression in normal tissues and tumor tissues at stage II and stage III. Inside lines represent means and standard deviations. **p,0.01. (B) Statistical results for the difference between normal and tumor tissues as analyzed by Kruskal-Wallis test. (C) Survival curves of breast cancer patients with respect to different SELENBP1 expression levels are shown at stage II and (D) stage III. Blue and red lines represent the SELENBP1-high and SELENBP1-low groups, respectively. doi:10.1371/journal.pone.0063702.g002

Article Snippet: Breast Cancer Tissue Array Slides Three sets of breast cancer tissue arrays (including normal, primary tumor, and metastases) were obtained from Imgenex or Biomax Inc.

Techniques: Expressing

Journal: PloS one

Article Title: Reduced selenium-binding protein 1 in breast cancer correlates with poor survival and resistance to the anti-proliferative effects of selenium.

doi: 10.1371/journal.pone.0063702

Figure Lengend Snippet: Figure 3. The Correlation of SELENBP1 Expression with ER, PR, and TP53 in Breast Cancer Tissues. (A) The scoring distributions of SELENBP1 expression in normal tissues and tumor tissues with ER+ and ER– status. The inside lines represent means and standard deviations. **p,0.01. The difference between normal and ER+ and ER– tumor tissues was analyzed by Kruskal-Wallis test and statistical results are shown (B). Survival curves of breast cancer patients with respect to different SELENBP1 expression are shown in ER+ group in (C). The blue line is the SELENBP1- high group and the red line is the SELENBP1-low group. The scoring distributions of SELENBP1 expression in normal and tumor tissues with PR+/PR–

Article Snippet: Breast Cancer Tissue Array Slides Three sets of breast cancer tissue arrays (including normal, primary tumor, and metastases) were obtained from Imgenex or Biomax Inc.

Techniques: Expressing

Journal: Molecular Vision

Article Title: Quantification of retinal pigment epithelial phenotypic variation using laser scanning cytometry

doi:

Figure Lengend Snippet: Primary and secondary antibodies used in this study.

Article Snippet: Slides were washed and incubated with a biotinylated secondary antibody (Vector Laboratories, Burlingame, CA) as per the manufactures instructions, washed, and incubated with the Vectastain ABC Reagent containing the enzyme alkaline phosphatase (Vector Laboratories), and washed once more with PBS before incubating with nitroblue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (NBT/BCIP; Vector Laboratories).

Techniques: Plasmid Preparation

Journal: Molecular Vision

Article Title: Quantification of retinal pigment epithelial phenotypic variation using laser scanning cytometry

doi:

Figure Lengend Snippet: Immunolabeling for Mn-superoxide dismutase (MnSOD) in human retinal pigment epithelium (RPE). Panel A shows a negative control section. Panel B shows a section from the macula of a 64-year-old female; the section was processed for immunohistochemistry with 4 μg/ml goat antibody against Mn-superoxide dismutase protein (anti-SOD2) primary antibody, 7.5 μg/ml biotinylated rabbit antigoat IgG, avidin/biotin complexed with alkaline phosphatase, and NBT/BCIP substrate. Arrow heads point to cone photoreceptors, and arrows point to RPE cells. The magnification bar represents 10 µ (400×). Abbreviations: NBT represents 4-Nitro blue tetrazolium, BCIP represents 5-bromo-4-chloro-3-indolyl-phosphate.

Article Snippet: Slides were washed and incubated with a biotinylated secondary antibody (Vector Laboratories, Burlingame, CA) as per the manufactures instructions, washed, and incubated with the Vectastain ABC Reagent containing the enzyme alkaline phosphatase (Vector Laboratories), and washed once more with PBS before incubating with nitroblue tetrazolium/5-bromo-4-chloro-3-indolyl phosphate (NBT/BCIP; Vector Laboratories).

Techniques: Immunolabeling, Negative Control, Immunohistochemistry, Avidin-Biotin Assay

Journal: bioRxiv

Article Title: Global kinome silencing combined with 3D invasion screening of the tumor microenvironment identifies fibroblast-expressed PIK3Cδ involvement in triple-negative breast cancer progression

doi: 10.1101/822049

Figure Lengend Snippet: ( A ) Schematic representation of the in vivo experiment using NOD CB17 PRKDC/J mice. MDA-MB-231 (Groups 1-2) and MDA-MB-231/MRC5 (Groups 3-4) tumor cells were implanted s.c. on day 0. After randomization on day 7, treatment with CAL-101 was initiated in Groups 2 and 4, whereas Groups 1 and 3 were administrated with vehicle. During the course of the study, the growth of the subcutaneously implanted primary tumors was determined twice weekly by luminescence and caliper measurement. ( B ) Upper panel: Box and whisker plots comparing different groups at Day14 and Day 21. Significance was calculated using unpaired t-test. Results are expressed as mean ± SEM; ** P < 0.01, ns: not significant. Lower panel: Representative in vivo images of different groups, treated with vehicle or CAL-101. ( D ) Histological analysis of Ki67 expression in representative tumor tissue sections of different groups. Original magnification, 20×. Scale bar, 50μm. ( D ) Representative images of immunofluorescent staining of MDA-MB-231/MRC5 tumor cryosections for α-SMA and p-AKT (Ser473) after vehicle or CAL-101 treatment. Significance was calculated using unpaired t-test. Results are expressed as mean ± SEM; * P < 0.05 vs. vehicle treated tumors. Original magnification, 40×. Scale bar, 50 μm. ( E ) Representative images of immunofluorescence staining of tumor cryosections using TE-7 anti-human fibroblast antibody. Original magnification, 20×. Scale bar, 100 μm.

Article Snippet: Slides were incubated overnight at 4 °C with antibodies to the following: α-SMA (Abcam, ab21027, 1:00), F4/80 (Biolegend, 123101, 1:20), PIK3Cδ (Santa Cruz, sc-55589, 1:30), pAkt-S473 (Cell Signaling Technology, CST, 4060, 1:100), pAkt-Thr308 (Millipore, 07-1398, 1:100), E-cadherin (CST, 14472S, 1:100), Vimentin (CST, 5741S, 1:100), CD31 (Abcam, ab9498, 1:100), Ki67 (Abcam, ab15580, 1:50).

Techniques: In Vivo, Whisker Assay, Expressing, Staining, Immunofluorescence

Journal: bioRxiv

Article Title: Global kinome silencing combined with 3D invasion screening of the tumor microenvironment identifies fibroblast-expressed PIK3Cδ involvement in triple-negative breast cancer progression

doi: 10.1101/822049

Figure Lengend Snippet: ( A ) Tumor volumes chart from MMTV-PyMT transgenic mice after vehicle or CAL-101 treatment (n=8 mice / group). Individual values for each mouse are displayed. Significance was calculated using unpaired t-test (week 12). Results are expressed as mean ± SEM; *** P < 0.001. (B) Representative images for immunohistochemical Ki-67 staining in the mammary tumor sections of MMTV-PyMT transgenic mice after vehicle or CAL-101 treatment. (C) Quantification of lung metastatic nodules in each group. Significance was calculated using unpaired t-test. Results are expressed as mean ± SEM; * P < 0.05. Yellow and black dots represent mice that were sacrificed at week 12 or week 15 respectively. (D) Left panel: Representative images of immunofluorescent staining for α-SMA and p-AKT (Thr308) in the mammary tumor sections of MMTV-PyMT transgenic mice after vehicle or CAL-101 treatment. Arrows indicate a-SMA + fibroblasts. Higher-magnification images are shown at the bottom right corner. Right panel: Quantification of p-AKT (Thr308) immunofluorescent staining in tumor infiltrating a-SMA + fibroblasts in the mammary tumors of MMTV-PyMT transgenic mice after vehicle or CAL-101 treatment. Significance was calculated using multiple t-tests. Results are expressed as mean ± SEM; ** P < 0.01 vs. vehicle treated tumors. ( E ) Left panel: Representative images of immunofluorescent staining for F4/80 and p-AKT (Thr308) in the mammary tumor sections of MMTV-PyMT transgenic mice after vehicle or CAL-101 treatment. Arrows indicate F4/80 + macrophages. Higher-magnification images are shown at the bottom right corner. Right panel: Quantification of p-AKT (Thr308) immunofluorescent staining in tumor infiltrating F4/80 + macrophages in the mammary tumors of MMTV-PyMT transgenic mice after vehicle or CAL-101 treatment. Significance was calculated using multiple t-tests. Results are expressed as mean ± SEM; * P < 0.05 vs. vehicle treated tumors. ( F ) Representative images of IHC staining of the mammary tumor sections of MMTV-PyMT transgenic mice stained for BDNF, PLGF and NR4A1, after vehicle or CAL-101 treatment. Significance was calculated using unpaired t-test. Results are expressed as mean ± SEM; * P < 0.05, *** P < 0.001 vs. vehicle treated tumors. Original magnification, 20×. Scale bar, 100 μm. The intensity of BDNF, PLGF and NR4A1 signals were evaluated in the vehicle treated tumor slides and then this value was compared with the signals from CAL-101 treated tumor slides results are expressed as percentage (%) of signal vs vehicle treated tumors.

Article Snippet: Slides were incubated overnight at 4 °C with antibodies to the following: α-SMA (Abcam, ab21027, 1:00), F4/80 (Biolegend, 123101, 1:20), PIK3Cδ (Santa Cruz, sc-55589, 1:30), pAkt-S473 (Cell Signaling Technology, CST, 4060, 1:100), pAkt-Thr308 (Millipore, 07-1398, 1:100), E-cadherin (CST, 14472S, 1:100), Vimentin (CST, 5741S, 1:100), CD31 (Abcam, ab9498, 1:100), Ki67 (Abcam, ab15580, 1:50).

Techniques: Transgenic Assay, Immunohistochemical staining, Staining, Immunohistochemistry

Journal: Journal of proteomics

Article Title: Plasma GPI and PGD are associated with vascular normalization and may serve as novel prognostic biomarkers for lung adenocarcinoma: Multi-omics and multi-dimensional analysis.

doi: 10.1016/j.jprot.2024.105247

Figure Lengend Snippet: Fig. 2. Screening and validation of candidate plasma proteins. (A) 1331 identified proteins in plasma, CP: cytoplasm; ER: endoplasmic reticulum; EC: extracellular; MC: mitochondria; PM: plasma membrane. (B) The differentially expressed proteins between the case and control groups are described using volcano plots. The differentially expressed proteins on the statistical criteria of fold change <2/3, >3/2 (log2 scaled), and P < 0.05 (−log10 scaled). (C) Differential expression of GPI between 102 LUAD cases and 102 healthy controls in the validation phase. (D) Differential expression of PGD between 102 LUAD cases and 102 healthy controls in the validation phase. Navy blue, total LUAD cases, red, total healthy controls; green, female LUAD cases, light blue, female healthy controls; purple, male LUAD cases, orange, male healthy controls. (E) ROC curve for GPI expression levels of LUAD cases versus healthy controls. (F) ROC curve for PGD expression levels of LUAD cases versus healthy controls. (G) ROC curve for combined GPI and PGD expression levels of LUAD cases versus healthy controls. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: For immunohistochemistry stainings, the slides were incubated with GPI (CST, 94068 T) or PGD (Proteintech, 14,718–1-AP) antibody solution for 24 h at 4 ◦C with gentle shaking.

Techniques: Biomarker Discovery, Clinical Proteomics, Membrane, Control, Quantitative Proteomics, Expressing

Journal: Journal of proteomics

Article Title: Plasma GPI and PGD are associated with vascular normalization and may serve as novel prognostic biomarkers for lung adenocarcinoma: Multi-omics and multi-dimensional analysis.

doi: 10.1016/j.jprot.2024.105247

Figure Lengend Snippet: Fig. 3. GPI and PGD are consistently differential expressed in both plasma and tissues. (A) GPI protein are differential expressed in plasma. (B) GPI protein are differential expressed in tissue. (C) GPI mRNA are differential expressed in tissue. (D) PGD protein are differential expressed in plasma. (E) PGD protein are dif ferential expressed in tissue. (F) PGD mRNA are differential expressed in tissue.

Article Snippet: For immunohistochemistry stainings, the slides were incubated with GPI (CST, 94068 T) or PGD (Proteintech, 14,718–1-AP) antibody solution for 24 h at 4 ◦C with gentle shaking.

Techniques: Clinical Proteomics

Journal: Journal of proteomics

Article Title: Plasma GPI and PGD are associated with vascular normalization and may serve as novel prognostic biomarkers for lung adenocarcinoma: Multi-omics and multi-dimensional analysis.

doi: 10.1016/j.jprot.2024.105247

Figure Lengend Snippet: Fig. 4. Kaplan-Meier estimates the overall survival probability of LUAD patients. (A-B) Kaplan-Meier survival curve analysis of the prognostic significance of a high and a low mRNA expression of GPI and PGD. (C–D) Kaplan-Meier survival curve analysis of the prognostic significance of a high and a low protein expression of GPI and PGD.

Article Snippet: For immunohistochemistry stainings, the slides were incubated with GPI (CST, 94068 T) or PGD (Proteintech, 14,718–1-AP) antibody solution for 24 h at 4 ◦C with gentle shaking.

Techniques: Expressing

Journal: Journal of proteomics

Article Title: Plasma GPI and PGD are associated with vascular normalization and may serve as novel prognostic biomarkers for lung adenocarcinoma: Multi-omics and multi-dimensional analysis.

doi: 10.1016/j.jprot.2024.105247

Figure Lengend Snippet: Fig. 6. Association between expression of GPI and PGD in tissue and plasma and vascular normalization (**P < 0.01; ***P < 0.001; ****P < 0.0001).

Article Snippet: For immunohistochemistry stainings, the slides were incubated with GPI (CST, 94068 T) or PGD (Proteintech, 14,718–1-AP) antibody solution for 24 h at 4 ◦C with gentle shaking.

Techniques: Expressing, Clinical Proteomics

Journal: Journal of proteomics

Article Title: Plasma GPI and PGD are associated with vascular normalization and may serve as novel prognostic biomarkers for lung adenocarcinoma: Multi-omics and multi-dimensional analysis.

doi: 10.1016/j.jprot.2024.105247

Figure Lengend Snippet: Fig. 5. GO functional classification and KEGG pathway mapping of GPI, PGD, and relevant genes. (A) The STRING database was used to analyze the top 50 relevant genes and Cytoscape was used to display the protein-protein interaction (PPI) network. (B) The distributions were summarized in three main categories: biological process, molecular function (MF), and cellular component (CC). (C) Scatter plot of enriched KEGG pathways statistics. The color and size of the dots represent the range of the P-value and the number of genes mapped to the indicated pathways, respectively. Top 10 enriched pathways are shown in the fig. (D) The map of “glycolysis” was modified from the KEGG map.

Article Snippet: For immunohistochemistry stainings, the slides were incubated with GPI (CST, 94068 T) or PGD (Proteintech, 14,718–1-AP) antibody solution for 24 h at 4 ◦C with gentle shaking.

Techniques: Functional Assay, Modification

Journal: Journal of proteomics

Article Title: Plasma GPI and PGD are associated with vascular normalization and may serve as novel prognostic biomarkers for lung adenocarcinoma: Multi-omics and multi-dimensional analysis.

doi: 10.1016/j.jprot.2024.105247

Figure Lengend Snippet: Fig. 7. Lung adenocarcinoma active metastasis model. (A) Schematic of active metastasis tumor-bearing mice model, tumor tissues of mice without lung metastasis (a), the tumor tissues of mice with lung metastasis (b) and the corresponding lung metastasis tumor tissues (c). (B) Immunofluorescence photographs of tumor tissue of mice without lung metastasis (a), the tumor tissues of mice with lung metastasis (b) with CD31 and α-SMA staining. Scale: 200 μM. (C) Differential expression of α-SMA in tumor tissue of mice without lung metastasis (a), the tumor tissues of mice with lung metastasis (b). (D) The expression of α-SMA, GPI, PGD in tumor tissues of mice without lung metastasis(a), the tumor tissues of mice with lung metastasis (b) and the corresponding lung metastasis tumor tissues (c). (E) Differential expression of α-SMA, GPI, and PGD in tumor tissue of mice without lung metastasis(a), the tumor tissues of mice with lung metastasis (b) and the corresponding lung metastasis tumor tissues (c). (*P < 0.05; ***P < 0.001).

Article Snippet: For immunohistochemistry stainings, the slides were incubated with GPI (CST, 94068 T) or PGD (Proteintech, 14,718–1-AP) antibody solution for 24 h at 4 ◦C with gentle shaking.

Techniques: Immunofluorescence, Staining, Quantitative Proteomics, Expressing