Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: Confocal microscopy (lung, liver, spleen, brain) of PKH67-labeled Panc02 EXO and Panc02-H7 EXO tissue distribution (green) 24 hpi. (A) PKH-67-labeled liposomes served as controls (scale bar=100 μm). Histogram shows exosome tissue distribution quantification (n=5/group). CD45, p-Stat3, and CD11b IF staining in liver sections from controls (left) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (right) for 12 d without tumor challenge. (B) Histogram shows infiltrating CD45 + cell quantification. FN and α-SMA IF staining in liver sections from controls (top) and mice treated with Panc02 EXOs(middle) or Panc02-H7 EXOs (bottom) for 12 d without tumor challenge. (C) Histogram shows infiltrating α-SMA + hStCs and FN expression quantification(400× magnification; n=5/group). Western blotting analysis showed upregulated S100A8 and S100A9 in livers treated with Panc02-H7-derived exosomes. Histogram shows expression of the three proteins in three groupsas determined by densitometric analysis (n=3/group). (D) Pancreatic cancer-derived exosomes induce MDSC accumulation in peripheral blood. (E) Representative flow cytometric plots (left) and quantification (right) of CD11b + GR1 + MDSCs (n=5/group). *P<0.05, **P<0.01,***P<0.001.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques: Confocal Microscopy, Labeling, Liposomes, Staining, Expressing, Western Blot, Derivative Assay

Journal: Oncotarget

Article Title: Pancreatic cancer-derived exosomes promote tumor metastasis and liver pre-metastatic niche formation

doi: 10.18632/oncotarget.18831

Figure Lengend Snippet: IHC analysis and histopathological examination of macrophages (F4/80), hStCs (α-SMA), and neutrophils in liver metastatic niches of naïve mice and mice treated with PBS, Panc02 EXOs, or Panc02-H7 EXOs at 30d post-SOI (arrow shows neutrophils in liver) (A) . Representative histogram shows quantification of F4/80 + macrophages, α-SMA + hStCs, and neutrophils (B) . Identification of FN, S100A8, and S100A9 as inflammatory mediators, and collagen deposition in the liver metastatic niche (C) . Representative histogram shows FN and MTS quantification (D) . Representative histogram shows S100A8 and S100A9 quantification (E) . n=6/group.**P<0.01,***P<0.001.10 fields assessed per sample. FOV, field of view.

Article Snippet: Primary antibodies against fibronectin (1:100),α-SMA (1:50), S100A8 (1:50), S100A9 (1:50), and F4/80 (1:50) were purchased from Proteintech (Wuhan, China).

Techniques:

Journal: Frontiers in Oncology

Article Title: Identification of a Two-Gene ( PML-EPB41 ) Signature With Independent Prognostic Value in Osteosarcoma

doi: 10.3389/fonc.2019.01578

Figure Lengend Snippet: The functional analysis of candidate genes. (A) Visualization for functional enrichment results of 733 candidate genes by Cytoscape software. Each node represents an annotation term. (B) The results of g: profiler functional enrichment with prognostic candidate genes. Horizontal axis above represents annotation terms. Vertical axis represents the value of –log 10 ( p -value). (C) The Kaplan–Meier curves for overall survival of high- and low-risk groups of OSA in the internal training dataset and external validating dataset. The prognostic differences between the two groups in internal training dataset was significant (log-rank test p = 0.0057). (D) ROC of PML-EPB41 signature based on PI in training dataset samples. (E) ROC of PML signature in training dataset samples. (F) ROC of EPB41 signature in training dataset samples.

Article Snippet: The EPB41 antibody (Cat. no. 13014-1-AP), PML antibody (Cat. no. 21041-1-AP) and GAPDH antibody (Cat. no. 60004-1-Ig) were obtained from Proteintech.

Techniques: Functional Assay, Software

Journal: Frontiers in Oncology

Article Title: Identification of a Two-Gene ( PML-EPB41 ) Signature With Independent Prognostic Value in Osteosarcoma

doi: 10.3389/fonc.2019.01578

Figure Lengend Snippet: Result of univariate Cox regression analysis.

Article Snippet: The EPB41 antibody (Cat. no. 13014-1-AP), PML antibody (Cat. no. 21041-1-AP) and GAPDH antibody (Cat. no. 60004-1-Ig) were obtained from Proteintech.

Techniques:

Journal: Frontiers in Oncology

Article Title: Identification of a Two-Gene ( PML-EPB41 ) Signature With Independent Prognostic Value in Osteosarcoma

doi: 10.3389/fonc.2019.01578

Figure Lengend Snippet: Result of multivariate Cox regression analysis.

Article Snippet: The EPB41 antibody (Cat. no. 13014-1-AP), PML antibody (Cat. no. 21041-1-AP) and GAPDH antibody (Cat. no. 60004-1-Ig) were obtained from Proteintech.

Techniques:

Journal: Frontiers in Oncology

Article Title: Identification of a Two-Gene ( PML-EPB41 ) Signature With Independent Prognostic Value in Osteosarcoma

doi: 10.3389/fonc.2019.01578

Figure Lengend Snippet: (A) Relative mRNA expression level of PML and EPB41 in three cell lines are conducted by quantitative polymerase chain reaction. Data are presented as the mean ± SD. * P < 0.05 vs. hFOB1.19. (B) Expression of EPB41 and PML by western blot. Protein levels were statistically evaluated in columns. Measurements were in triplicate and data were presented as the mean ± SD. ** P < 0.01 vs. hFOB1.19.

Article Snippet: The EPB41 antibody (Cat. no. 13014-1-AP), PML antibody (Cat. no. 21041-1-AP) and GAPDH antibody (Cat. no. 60004-1-Ig) were obtained from Proteintech.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Cell Reports Medicine

Article Title: Alarmin S100A8 imparts chemoresistance of esophageal cancer by reprogramming cancer-associated fibroblasts

doi: 10.1016/j.xcrm.2024.101576

Figure Lengend Snippet:

Article Snippet: Human S100A8/S100A9 ELISA Kit , Proteintech , Cat# KE00177.

Techniques: Virus, shRNA, Over Expression, Recombinant, Purification, Staining, Enzyme-linked Immunosorbent Assay, Sonication, Chromatin Immunoprecipitation, Software

Journal: NPJ Vaccines

Article Title: Protective efficacy of the pan-fungal vaccine NXT-2 against vulvovaginal candidiasis in a murine model

doi: 10.1038/s41541-025-01171-4

Figure Lengend Snippet: IL-1β ( A ) and S100A8 ( B ) concentrations were assessed via ELISA. All data represents mean ± SEM. Statistical significance was assessed by using the Mann-Whitney test for IL-1β and S100A8 concentrations. C Tissue damage was assessed by measuring lactate dehydrogenase (LDH) activity in vaginal lavage fluid twenty-eight days post vaccination (28dpv) and five days post challenge (5dpc). Data represents mean ± SEM. Statistical significance was assessed by two-way ANOVA with Tukey correction (* P = 0.0111, *** P = 0.0007, **** P < 0.0001). D Quantification of inflammation in vaginal tissue from sham and NXT-2 immunized mice excised 5 days post challenge. Data represents mean ± SEM. Statistical significance was assessed by unpaired student t -test (**** P < 0.0001). Vaginal tissue excised following immunization and C. albicans challenge was stained with H&E (sham— E , NXT-2— G ) and the neutrophil specific marker anti-mouse Ly6G (sham— F , NXT-2— H ). Tissue sections were scanned at 40× objective magnification using the Aperio AT2 (Leica Biosystems). Image analysis and capture was performed using ImageScope (Leica Biosystems) at 20× digital zoom level.

Article Snippet: S100A8 : Utilizing the Mouse S100A8 ELISA kit (Rockland Immunochemicals), samples were diluted 1:100 and the manufacturer’s instructions were followed.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Activity Assay, Staining, Marker

Journal: International journal of molecular sciences

Article Title: The Role of Erythrocyte Membrane Protein Band 4.1-like 3 in Idiopathic Pulmonary Fibrosis.

doi: 10.3390/ijms241210182

Figure Lengend Snippet: Figure 1. Comparison of mRNA and protein EPB41L3 levels in lung-tissue-derived fibroblasts between 14 IPF patients and 10 controls. Expression of EPB41L3 mRNA (normalized to that of β-actin) was measured using (A) RT-PCR, (B) densitometry, and (C) real-time PCR (log (2−∆∆Ct). Protein expression of EPB41L3 (normalized to that of β-actin) was measured using (D) Western blot and (E) densitometry. The Mann–Whitney U test was performed to identify the statistical significance between IPF and control groups. Data are medians and quantiles, and * p < 0.01. (F) Correlations between the RT-PCR and Western blot band intensities was analyzed using Spearman’s correlation coefficient. (See Figure S1 for RT-PCR and original Western blot images.)

Article Snippet: The membranes were blocked in 5% skimmed milk and incubated for 24 h at 4 ◦C with the following primary antibodies: rabbit polyclonal anti-human EPB41L3 (1:2000; ProteinTech, Rosemont, IL, USA, #10719-I-AP), mouse monoclonal anti-human E-cadherin (1:1000; Invitrogen, #33-4000), mouse monoclonal anti-human N-cadherin (1:1000, Invitrogen, #33-3300), rabbit polyclonal anti-human collagen I (1:1000; Abcam, Cambridge, MA, USA, #MA1-26771), mouse monoclonal anti-human-α-SMA (1:500; Abcam, #ab7817), mouse monoclonal anti-human fibronectin (1:1000; Abcam, #ab6328), and mouse monoclonal anti-human β-actin (1:50,000; Sigma-Aldrich, St. Louis, MO, USA, #A1978).

Techniques: Comparison, Derivative Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, MANN-WHITNEY, Control

Journal: International journal of molecular sciences

Article Title: The Role of Erythrocyte Membrane Protein Band 4.1-like 3 in Idiopathic Pulmonary Fibrosis.

doi: 10.3390/ijms241210182

Figure Lengend Snippet: Figure 2. Changes in mRNA and protein of EPB41L3 and EMT-related genes expressed by A549 cells after stimulation with TGF-β. A549 cells were cultured with or without 5 ng/mL of TGF-β1 for 24, 48, and 72 h. The EPB41L3-, N-cadherin-, E-cadherin-, and COL1A1-to-β-actin ratios were measured using (A) real-time PCR, (B) Western blot, and (C) densitometry of protein bands. They were normalized to β-actin and expressed as ratios. All experiments were performed separately six times, and the representative images are shown. Data are mean ± SE of the six independent experiments. Basal levels (0 h) were excluded from statistical analysis. Significant differences between groups were evaluated using two-way ANOVA with Bonferroni multiple comparisons test. * p < 0.05 and ** p < 0.01 compared to TGF-β1-stimulated cells. Black bars: untreated control group, bars with diagonals: TGF-β1-treated group. (See Figure S2 for original Western blot images).

Article Snippet: The membranes were blocked in 5% skimmed milk and incubated for 24 h at 4 ◦C with the following primary antibodies: rabbit polyclonal anti-human EPB41L3 (1:2000; ProteinTech, Rosemont, IL, USA, #10719-I-AP), mouse monoclonal anti-human E-cadherin (1:1000; Invitrogen, #33-4000), mouse monoclonal anti-human N-cadherin (1:1000, Invitrogen, #33-3300), rabbit polyclonal anti-human collagen I (1:1000; Abcam, Cambridge, MA, USA, #MA1-26771), mouse monoclonal anti-human-α-SMA (1:500; Abcam, #ab7817), mouse monoclonal anti-human fibronectin (1:1000; Abcam, #ab6328), and mouse monoclonal anti-human β-actin (1:50,000; Sigma-Aldrich, St. Louis, MO, USA, #A1978).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International journal of molecular sciences

Article Title: The Role of Erythrocyte Membrane Protein Band 4.1-like 3 in Idiopathic Pulmonary Fibrosis.

doi: 10.3390/ijms241210182

Figure Lengend Snippet: Figure 3. Effect of EPB41L3 transfection on EMT-related genes in the A549 cell line. Changes in EPB41L3 and EMT-related gene expression after EPB41L3 transfection in the presence and absence of 5 ng/mL of TGF-β1 for 48 h. Expression of mRNA and protein normalized to β-actin levels were measured using (A-1–A-4) real-time PCR, (B) Western blot, and (C-1–C-4) densitometry of protein bands. They were normalized to β-actin and expressed as ratios. Independent experiments were analyzed using densitometry. All experiments were performed separately six times, and the represen- tative images are shown. Data are mean ± SE of 6 independent experiments. Significant differences between groups were evaluated using two-way ANOVA with Bonferroni multiple comparisons test. * p < 0.05 and ** p < 0.01. EMT: epithelial–mesenchymal transition, Lv: Lentivirus, black bar: control group, open bar: EPB41L3-overexpressing group. (See Figure S3 for original Western blot images).

Article Snippet: The membranes were blocked in 5% skimmed milk and incubated for 24 h at 4 ◦C with the following primary antibodies: rabbit polyclonal anti-human EPB41L3 (1:2000; ProteinTech, Rosemont, IL, USA, #10719-I-AP), mouse monoclonal anti-human E-cadherin (1:1000; Invitrogen, #33-4000), mouse monoclonal anti-human N-cadherin (1:1000, Invitrogen, #33-3300), rabbit polyclonal anti-human collagen I (1:1000; Abcam, Cambridge, MA, USA, #MA1-26771), mouse monoclonal anti-human-α-SMA (1:500; Abcam, #ab7817), mouse monoclonal anti-human fibronectin (1:1000; Abcam, #ab6328), and mouse monoclonal anti-human β-actin (1:50,000; Sigma-Aldrich, St. Louis, MO, USA, #A1978).

Techniques: Transfection, Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International journal of molecular sciences

Article Title: The Role of Erythrocyte Membrane Protein Band 4.1-like 3 in Idiopathic Pulmonary Fibrosis.

doi: 10.3390/ijms241210182

Figure Lengend Snippet: Figure 4. Effect of silencing EPB41L3 on EMT-related genes in A549 cells. Changes of EPB41L3 expression in response to EPB41L3 siRNA in the presence and absence of 5 ng/mL of TGF-β1 for 48 h. Expression of mRNA and protein normalized to the β-actin level was measured using (A) real-time PCR, (B) Western blot, and (C-1–C-3) densitometry of protein bands. Independent experiments were analyzed using densitometry. They were normalized to β-actin and expressed as ratios. All experiments were performed separately six times, and the representative images are shown. Data are mean ± SE of 6 independent experiments. Significant differences between groups were evaluated using two-way ANOVA with Bonferroni multiple comparisons test. * p < 0.05. Black bar: control group, open bar: scramble group, Gray bar: EPB41L3-knockdown group. (See Figure S4 for original Western blot images).

Article Snippet: The membranes were blocked in 5% skimmed milk and incubated for 24 h at 4 ◦C with the following primary antibodies: rabbit polyclonal anti-human EPB41L3 (1:2000; ProteinTech, Rosemont, IL, USA, #10719-I-AP), mouse monoclonal anti-human E-cadherin (1:1000; Invitrogen, #33-4000), mouse monoclonal anti-human N-cadherin (1:1000, Invitrogen, #33-3300), rabbit polyclonal anti-human collagen I (1:1000; Abcam, Cambridge, MA, USA, #MA1-26771), mouse monoclonal anti-human-α-SMA (1:500; Abcam, #ab7817), mouse monoclonal anti-human fibronectin (1:1000; Abcam, #ab6328), and mouse monoclonal anti-human β-actin (1:50,000; Sigma-Aldrich, St. Louis, MO, USA, #A1978).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Knockdown

Journal: International journal of molecular sciences

Article Title: The Role of Erythrocyte Membrane Protein Band 4.1-like 3 in Idiopathic Pulmonary Fibrosis.

doi: 10.3390/ijms241210182

Figure Lengend Snippet: Figure 5. Changes in mRNA and protein levels of EPB41L3 and FMT-related genes expressed by MRC5 cells after stimulation with TGF-β. MRC5 cells were cultured with or without 5 ng/mL of TGF-β1 for 24, 48, and 72 h. The EPB41L3-, FN1-, ACTA2-, N-cadherin-, and COL1A1-to-β-actin ratios were measured using (A-1–A-5) real-time PCR, (B) Western blot and (C-1–C-4) densitometry. Independent experiments were analyzed using densitometry. They were normalized to β-actin and expressed as ratios. All experiments were performed separately six times, and the representative images are shown. Data are mean ± SE of 6 independent experiments. Basal levels (0 h) were excluded from statistical analysis. Significant differences between groups were evaluated using two-way ANOVA with Bonferroni multiple comparisons test. * p < 0.05, ** p < 0.01 and *** p < 0.001 compared to TGF-β1-stimulated cells. Black bar: control group, open bar: EPB41L3-overexpressing group. (See Figure S5 for original Western blot images).

Article Snippet: The membranes were blocked in 5% skimmed milk and incubated for 24 h at 4 ◦C with the following primary antibodies: rabbit polyclonal anti-human EPB41L3 (1:2000; ProteinTech, Rosemont, IL, USA, #10719-I-AP), mouse monoclonal anti-human E-cadherin (1:1000; Invitrogen, #33-4000), mouse monoclonal anti-human N-cadherin (1:1000, Invitrogen, #33-3300), rabbit polyclonal anti-human collagen I (1:1000; Abcam, Cambridge, MA, USA, #MA1-26771), mouse monoclonal anti-human-α-SMA (1:500; Abcam, #ab7817), mouse monoclonal anti-human fibronectin (1:1000; Abcam, #ab6328), and mouse monoclonal anti-human β-actin (1:50,000; Sigma-Aldrich, St. Louis, MO, USA, #A1978).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International journal of molecular sciences

Article Title: The Role of Erythrocyte Membrane Protein Band 4.1-like 3 in Idiopathic Pulmonary Fibrosis.

doi: 10.3390/ijms241210182

Figure Lengend Snippet: Figure 6. Effect of EPB41L3 transfection on FMT-related genes in the MRC5 cell line. Changes in the expression of EPB41L3 and FMT-related genes in response to EPB41L3 transfection in the presence and absence of 5 ng/mL of TGF-β1 for 48 h. Expression of mRNA and protein normalized to the β-actin level were measured using (A-1–A-3) real-time PCR, (B) Western blot, and (C-1–C-3) densitometry of protein bands. They were normalized to β-actin and expressed as ratios. Independent experiments were analyzed using densitometry. All experiments separately were performed six times, and the representative images are shown. Data are mean ± SE of 6 independent experiments. Significant differences between groups were evaluated using two-way ANOVA with Bonferroni multiple comparisons test. * p < 0.05 and ** p < 0.01. Lv: Lentivirus, black bar: control group, open bar: EPB41L3-overexpressing group. (See Figure S6 for original Western blot images).

Article Snippet: The membranes were blocked in 5% skimmed milk and incubated for 24 h at 4 ◦C with the following primary antibodies: rabbit polyclonal anti-human EPB41L3 (1:2000; ProteinTech, Rosemont, IL, USA, #10719-I-AP), mouse monoclonal anti-human E-cadherin (1:1000; Invitrogen, #33-4000), mouse monoclonal anti-human N-cadherin (1:1000, Invitrogen, #33-3300), rabbit polyclonal anti-human collagen I (1:1000; Abcam, Cambridge, MA, USA, #MA1-26771), mouse monoclonal anti-human-α-SMA (1:500; Abcam, #ab7817), mouse monoclonal anti-human fibronectin (1:1000; Abcam, #ab6328), and mouse monoclonal anti-human β-actin (1:50,000; Sigma-Aldrich, St. Louis, MO, USA, #A1978).

Techniques: Transfection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: International journal of molecular sciences

Article Title: The Role of Erythrocyte Membrane Protein Band 4.1-like 3 in Idiopathic Pulmonary Fibrosis.

doi: 10.3390/ijms241210182

Figure Lengend Snippet: Figure 7. Effect of silencing EPB41L3 on FMT-related genes in MRC5 cells. Changes in the ex- pression of EPB41L3 and FMT-related genes in response to EPB41L3 siRNA. Expression of mRNA and protein were measured using (A-1–A-4) real-time PCR, (B) Western blot, and (C-1–C-4) den- sitometry of protein bands. Independent experiments were analyzed using densitometry. They were normalized to β-actin and expressed as ratios. All experiments separately were performed six times, and the representative images are shown. Data are mean ± SE of 6 independent experiments. Significant differences between groups were evaluated using two-way ANOVA with Bonferroni multiple comparisons test. * p < 0.05. Black bar: control group, open bar: scramble group, gray bar: EPB41L3-knockdown group. (See Figure S7 for original Western blot images).

Article Snippet: The membranes were blocked in 5% skimmed milk and incubated for 24 h at 4 ◦C with the following primary antibodies: rabbit polyclonal anti-human EPB41L3 (1:2000; ProteinTech, Rosemont, IL, USA, #10719-I-AP), mouse monoclonal anti-human E-cadherin (1:1000; Invitrogen, #33-4000), mouse monoclonal anti-human N-cadherin (1:1000, Invitrogen, #33-3300), rabbit polyclonal anti-human collagen I (1:1000; Abcam, Cambridge, MA, USA, #MA1-26771), mouse monoclonal anti-human-α-SMA (1:500; Abcam, #ab7817), mouse monoclonal anti-human fibronectin (1:1000; Abcam, #ab6328), and mouse monoclonal anti-human β-actin (1:50,000; Sigma-Aldrich, St. Louis, MO, USA, #A1978).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Knockdown