Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Flavokawain A alleviates the progression of mouse osteoarthritis: An in vitro and in vivo study

doi: 10.3389/fbioe.2022.1071776

Figure Lengend Snippet: Protective effects of FKA against IL-1β-induced inflammation and ECM destruction in mouse chondrocytes. Chondrocyte cell treatment with 5 ng/ml of IL-1β only or with FKA (20 and 40 μM) for a period of 24 h. Western blot analysis of inflammatory cytokines (COX2 and iNOS), catabolic (ADAMTS5, MMP3, and MMP13), and anabolic markers (Col2, Aggrecan, and Sox9) (A,C and E) . Quantitative analysis of protein expression (B,D and F) . (G and H) 5 ng/ml of IL-1β was added for treating the cells for 24 h in the absence or presence of 40 µM FKA. Confocal microscopy showing the expression of MMP13 and Aggrecan in immunofluorescence experiments (scale bar 10 μm). (I) Relative quantification of fluorescence intensity. The values are presented as means ± SD ( n = 3). # p < 0.05 vs the control group; * p < 0.05 and ** p < 0.01 vs the IL-1β group.

Article Snippet: ADAMTS5 primary antibody, secondary antibody, phosphate-buffered saline (PBS), trypsin, collagenase type II, the CCK8 assay kit, bovine serum albumin (BSA), and protein extraction kit were ordered and acquired from Boster Biological Technology (Wuhan, Hubei, China).

Techniques: Western Blot, Expressing, Confocal Microscopy, Immunofluorescence, Quantitative Proteomics, Fluorescence, Control

Journal: The Journal of pathology

Article Title: Single-cell profiling identifies distinct hormonal, immunologic, and inflammatory signatures of endometriosis-constituting cells.

doi: 10.1002/path.6178

Figure Lengend Snippet: Figure 4. Subpopulations and predicted interaction of myeloid cells. (A) UMAP plot showing 14 myeloid subpopulations identified (n = 7,159 cells, cDC1: type 1 conventional dendritic cell, cDC2: type 2 conventional dendritic cell, mPC: mature dendritic cell, pDC: plasmacytoid dendritic cell, Mo-Mac: monocyte-derived macrophage, ncMo: nonclassical monocyte, resMo: resident monocyte, Mac: macrophage). (B) Canonical gene markers for each myeloid subpopulation. (C) Overall proportions of 13 myeloid subpopulations (color legend) according to OEC, SPE, and DIE tissues. (D) UMAP plot colored by NE and endometriosis tissue. (E) Dot plot showing genes involved in proinflammatory reaction and immunotolerance in Mo-Mac subpopulation. (F) Predicted ligand-receptor interaction between Schwann cell and Mo-Mac subpopulations. Bubble plot showing NRG1-ERBB3 pair (upper). UMAP plot showing NRG1 expression in myeloid cells (middle left) and ERBB3 expression in Schwann cells (middle right). IHC showing positive ERBB3 expression in Schwann cells (N) and positive NRG1 expression in mononuclear cells around nerve (circle) of DIE. In comparison, ERBB3 expression in Schwann cells is weak (N) in adenomyosis, where the number of NRG1-expressing macrophages is low (arrows).

Article Snippet: We used antibodies against ARID1A (Abcam, Cambridge, UK, clone EPR13501; dilution 1/1000) [33], ERBB3 (Atlas Antibodies AB, Bromma, Sweden, clone HPA045396; 1/50), and NRG1 (Atlas Antibodies AB, clone HPA010964; 1/50).

Techniques: Derivative Assay, Expressing, Comparison

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Overexpression of ERBB3 promotes proliferation, migration, and angiogenesis in nasopharyngeal carcinoma

doi:

Figure Lengend Snippet: Expression levels of ERBB3 in NPC tissues and cell lines. A. Expression levels of ERBB3 in NPC tissues and normal nasopharyngeal epithelial tissues. Some representative results are shown. B, C. Protein levels of ERBB3 in 4 NPC tissues and 4 normal nasopharyngeal tissues by western blot analysis. *P < 0.05. **P < 0.01.

Article Snippet: The membrane was incubated with the anti-ERBB3 primary antibody (1:1000, Abclonal) or anti-VEGFA polyclonal antibody (1:300, BBI Life Sciences) overnight at 4°C and then incubated with HRP-tagged secondary antibodies (1:2000, BBI Life Sciences) at room temperature for 1 h. Immunoreactivity was detected by ECL reagent (Millipore) and quantitative data were obtained using Image J software.

Techniques: Expressing, Western Blot

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Overexpression of ERBB3 promotes proliferation, migration, and angiogenesis in nasopharyngeal carcinoma

doi:

Figure Lengend Snippet: Knockdown of ERBB3 inhibited the proliferation of CNE-2 cells. A, B. Western blot showed the expression of ERBB3 after CNE-2 cells transfected with ERBB3-specific siRNAs and a negative control siRNA. C. Proliferation was detected by CCK-8 assay after CNE-2 cells were treated with ERBB3-siRNAs or negative control siRNA for the indicated time. D. Histogram shows CNE-2 cell survival at 0 h and 60 h. E. Cell cycle analysis by flow cytometry in CNE-2 with ERBB3 downregulation. The data shown represent at least three independent experiments. *P < 0.05. **P < 0.01.

Article Snippet: The membrane was incubated with the anti-ERBB3 primary antibody (1:1000, Abclonal) or anti-VEGFA polyclonal antibody (1:300, BBI Life Sciences) overnight at 4°C and then incubated with HRP-tagged secondary antibodies (1:2000, BBI Life Sciences) at room temperature for 1 h. Immunoreactivity was detected by ECL reagent (Millipore) and quantitative data were obtained using Image J software.

Techniques: Knockdown, Western Blot, Expressing, Transfection, Negative Control, CCK-8 Assay, Cell Cycle Assay, Flow Cytometry

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Overexpression of ERBB3 promotes proliferation, migration, and angiogenesis in nasopharyngeal carcinoma

doi:

Figure Lengend Snippet: Suppression of ERBB3 inhibited the migration of CNE-2 cells. A. Transwell assay used to detect the penetration of ERBB3-silencing cells through the membrane compared with controls. B. Absolute number of cells migrated through the membrane. C. Silenced and control cells were grown until confluence and their migratory capabilities were analyzed by wound-healing assay. Representative images of wound-healing assay are shown at 0 h and 90 h. D. Histogram shows the relative migration distance of cells. Migration speed of the cells was analyzed by the wound width/distance measured at 0 h. The data shown represent at least three independent experiments. *P < 0.05. **P < 0.01.

Article Snippet: The membrane was incubated with the anti-ERBB3 primary antibody (1:1000, Abclonal) or anti-VEGFA polyclonal antibody (1:300, BBI Life Sciences) overnight at 4°C and then incubated with HRP-tagged secondary antibodies (1:2000, BBI Life Sciences) at room temperature for 1 h. Immunoreactivity was detected by ECL reagent (Millipore) and quantitative data were obtained using Image J software.

Techniques: Migration, Transwell Assay, Membrane, Control, Wound Healing Assay

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Overexpression of ERBB3 promotes proliferation, migration, and angiogenesis in nasopharyngeal carcinoma

doi:

Figure Lengend Snippet: Knockdown of ERBB3 impacts the proliferation of HUVECs and the length of blood vessels. A, B. Western blot shows the expression of protein levels after knocking down ERBB3 in HUVECs. C. Cell proliferation was measured by CCK-8 assay after HUVECs were treated with ERBB3-siRNAs or negative control siRNA for the indicated time. D. The statistics of SIV length in 72 hpf Tg (fli1a: EGFP) embryos injected with negative control siRNA or ERBB3-siRNAs. E. Morphology of SIVs in 72 hpf Tg (fli1a: EGFP) embryos injected with negative control siRNA or ERBB3-siRNAs. The data shown represent at least three independent experiments. *P < 0.05. **P < 0.01.

Article Snippet: The membrane was incubated with the anti-ERBB3 primary antibody (1:1000, Abclonal) or anti-VEGFA polyclonal antibody (1:300, BBI Life Sciences) overnight at 4°C and then incubated with HRP-tagged secondary antibodies (1:2000, BBI Life Sciences) at room temperature for 1 h. Immunoreactivity was detected by ECL reagent (Millipore) and quantitative data were obtained using Image J software.

Techniques: Knockdown, Western Blot, Expressing, CCK-8 Assay, Negative Control, Injection

Journal: International Journal of Clinical and Experimental Pathology

Article Title: Overexpression of ERBB3 promotes proliferation, migration, and angiogenesis in nasopharyngeal carcinoma

doi:

Figure Lengend Snippet: Knockdown of ERBB3 reduced angiogenesis by downregulating vascular endothelial growth factor A (VEGF-A). A. Human VEGF-A Precoated ELISA Kit was used to measure serum VEGF-A levels in 5 NPC patients and 5 healthy controls. B, C. Western blot shows the expression of VEGF-A after knocking down ERBB3 in HUVECs. The data shown represent at least three independent experiments. *P < 0.05. **P < 0.01.

Article Snippet: The membrane was incubated with the anti-ERBB3 primary antibody (1:1000, Abclonal) or anti-VEGFA polyclonal antibody (1:300, BBI Life Sciences) overnight at 4°C and then incubated with HRP-tagged secondary antibodies (1:2000, BBI Life Sciences) at room temperature for 1 h. Immunoreactivity was detected by ECL reagent (Millipore) and quantitative data were obtained using Image J software.

Techniques: Knockdown, Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

Journal: Cell Death & Disease

Article Title: Anti-Trop2 blockade enhances the therapeutic efficacy of ErbB3 inhibition in head and neck squamous cell carcinoma

doi: 10.1038/s41419-017-0029-0

Figure Lengend Snippet: a The table shows the derivation of the models used in the study. b Growth curves of patient-derived xenografts treated with five weeks of anti-ErbB3 antibody. Tumors were allowed to grow to 150 mm 3 prior to treatment for five weeks. Forty-eight hours after the 5th week residual tumor nodules were harvested. n = 10 mice per group. Differences in growth between treated and untreated mice were significant to P < 0.0001 (ANOVA) in all experiments

Article Snippet: Mice were treated with intraperitoneal injections of anti-ErbB3 antibody (Genentech) at 25 milligram/kilogram (mg/kg) weekly, anti-Trop2 antibody (Pfizer) at 20 mg/kg weekly, the combination of the two at the same dose and schedule, or with PBS for negative control.

Techniques: Derivative Assay

Journal: Cell Death & Disease

Article Title: Anti-Trop2 blockade enhances the therapeutic efficacy of ErbB3 inhibition in head and neck squamous cell carcinoma

doi: 10.1038/s41419-017-0029-0

Figure Lengend Snippet: a Tumor cell lysates obtained from tumor nodules containing persister cells after 5 weeks of treatment of anti-ErbB3 antibody treatment shows reduced levels of p-EGFR (Y1068) in ErbB3-antibody-treated mice compared to vehicle treatment. b Representative immunoblots of protein lysates from conditionally reprogrammed cells (CRCs) from four PDX models show reduced levels of p-EGFR after six days of incubation with anti-ErbB3 antibody ( n = 3) or c three days after transient siRNA-mediated knockdown of ErbB3 ( n = 3) or d neuregulin-1 ( n = 3). Antibodies to both Y1068 and Y1045 were used. Relative decreases in phosphoprotein levels in control vs. experimental groups were quantified by photodensitometry after normalization to total EGFR or ErbB3. Squamous cells were separated from Swiss 3T3 feeder cells by differential trypsanization prior to preparation of protein lysates

Article Snippet: Mice were treated with intraperitoneal injections of anti-ErbB3 antibody (Genentech) at 25 milligram/kilogram (mg/kg) weekly, anti-Trop2 antibody (Pfizer) at 20 mg/kg weekly, the combination of the two at the same dose and schedule, or with PBS for negative control.

Techniques: Western Blot, Incubation, Knockdown, Control

Journal: Cell Death & Disease

Article Title: Anti-Trop2 blockade enhances the therapeutic efficacy of ErbB3 inhibition in head and neck squamous cell carcinoma

doi: 10.1038/s41419-017-0029-0

Figure Lengend Snippet: a Protein lysates taken from larger ( ≥ 12 mm, n = 10) compared to smaller ( ≤ 6 mm, n = 3) tumors show that EGFR and ErbB3 are co-upregulated coincident with HIF1α stabilization in larger tumors. b Protein lysates from conditionally reprogrammed cells (Hoc6) exposed to CoCl 2 show that two-hour pre-treatment with anti-ErbB3 antibody (left panel, n = 3) but not Cetuximab (right panel, n = 3) suppresses cobalt chloride-dependent HIF1α stabilization, which is maximal at three hours. Relative HIF1α levels in control vs. treatment groups were quantified by photodensitometry after normalization to actin

Article Snippet: Mice were treated with intraperitoneal injections of anti-ErbB3 antibody (Genentech) at 25 milligram/kilogram (mg/kg) weekly, anti-Trop2 antibody (Pfizer) at 20 mg/kg weekly, the combination of the two at the same dose and schedule, or with PBS for negative control.

Techniques: Control

Journal: Cell Death & Disease

Article Title: Anti-Trop2 blockade enhances the therapeutic efficacy of ErbB3 inhibition in head and neck squamous cell carcinoma

doi: 10.1038/s41419-017-0029-0

Figure Lengend Snippet: a Protein lysates from tumor nodules harvested from mice after 5 weeks of anti-ErbB3 antibody treatment reveals increased levels of Trop2 in persister cells. b Right panels are photomicrographs of Trop2 staining in FaDu tumor xenografts after five weeks of anti-ErbB3 antibody treatment; left panels show immunoblots from the same tumor xenografts. N = 5 per group. Trop2 levels were quantified by photodensitometry and normalized to actin. c Synergistic activity of ant-Trop2 and anti-ErbB3 antibodies against UW-SCC34 xenografts growing in mice. N = 5 per group

Article Snippet: Mice were treated with intraperitoneal injections of anti-ErbB3 antibody (Genentech) at 25 milligram/kilogram (mg/kg) weekly, anti-Trop2 antibody (Pfizer) at 20 mg/kg weekly, the combination of the two at the same dose and schedule, or with PBS for negative control.

Techniques: Staining, Western Blot, Activity Assay