Journal: Cell

Article Title: Serotonin reduction in post-acute sequelae of viral infection

doi: 10.1016/j.cell.2023.09.013

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Vil1 Taqman assay , Thermo Scientific , Mm00494146_m1.

Techniques: Virus, Clinical Proteomics, Recombinant, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Bicinchoninic Acid Protein Assay, SYBR Green Assay, TaqMan Assay, Software, Imaging, Control, Electron Microscopy, Low Protein Binding, Membrane

Journal: Immunity

Article Title: Lung γδ T cells mediate protective responses during neonatal influenza infection that are associated with Type-2 immunity

doi: 10.1016/j.immuni.2018.07.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The stained cells were resuspended in RNAse inhibitor containing sort buffer (RNAsin, Promega, 200U/μl) and sorted by gating on γδTCR + CD3 + cells directly into a 384-well PCR plate that had been preloaded with 1μl of reverse transcription reaction mix [1X SuperScript VILO Reaction Mix (Thermo Fisher), SuperScript VILO enzyme and 1X0.1% NP40 (Thermo Fisher)] with a sorter (Model SY3200, Sony Biotech Synergy sorter, Sony Biotech, San Jose, CA).

Techniques: Control, Virus, Plasmid Preparation, Recombinant, Cell Stimulation, Protease Inhibitor, Blocking Assay, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Bicinchoninic Acid Protein Assay, Sequencing, Software

Journal: Advanced Functional Materials

Article Title: Tumor‐On‐A‐Chip Model Incorporating Human‐Based Hydrogels for Easy Assessment of Metastatic Tumor Inter‐Heterogeneity

doi: 10.1002/adfm.202315940

Figure Lengend Snippet: Figure 5. Gene and protein biomarkers expression depending on tumor metastatic ability. a) Relative gene expression of VEGFA, COL1A1, MMP2, and MMP9 in the (i) low-metastatic and (ii) high-metastatic models for the different culture conditions, normalized to the static culture. b) Comparison of low- and high-metastatic relative gene expression of (i) VEGFA, (ii) COL1A1, and (iii) MMP2, for each culture condition. c) ELISA quantification of VEGFA protein secretion at days 3, 4, and 7. Statistical significance between different culture conditions, for the same time-point, is represented with a solid line. Unless otherwise stated, time-dependent significance in each culture condition and model is statistically different. d) Confocal images of Ezrin expression (green) in the low-metastatic model, at 7 days of culture. Actin filaments and nuclei are stained in red and blue, respectively. Scale bar: 200 μm. Data are presented as mean ± SD (n ≥3). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, ns: not significant.

Article Snippet: The samples were incubated with primary mouse monoclonal anti-human Ezrin antibody (1:50 in 5% FBS (v/v) in PBS, 3C12, Santa Cruz Biotechnology, USA) at 4 °C for 3 days, followed by PBS washing for 1 h, and then incubated with the secondary antibody goat antimouse Alexa Fluor 488 (1:100 in 5% FBS (v/v) in PBS, Thermo Fisher Scientific, USA) at 4 °C for 3 days.

Techniques: Expressing, Gene Expression, Comparison, Enzyme-linked Immunosorbent Assay, Staining

Journal: eLife

Article Title: Gq activity- and β-arrestin-1 scaffolding-mediated ADGRG2/CFTR coupling are required for male fertility

doi: 10.7554/eLife.33432

Figure Lengend Snippet: ( A ) Intracellular pH (pHi) of the ligated efferent ductules from WT (n = 9) mice and Adgrg2 -/Y (n = 9) mice were measured by carboxy-SNARF (5 μM), with or without incubation with the CFTR inhibitor CFTRinh-172. ( B ) qRT-PCR analysis of CFTR levels in the efferent ductules of WT (n = 3) or Adgrg2 -/Y (n = 3) mice. ( C ) Co-localization of ADGRG2 (red fluorescence) and CFTR (sc-8909, Santa Cruz, green fluorescence) in the male efferent ductules of WT mice. Scale bars, 50 μm. ( D ) Analysis of ADGRG2 and CFTR fluorescence intensities by Pearson’s correlation analysis. The Pearson's correlation coefficient was 0.76. ( E ) Immunofluorescence staining of ADGRG2 (red fluorescence) and CFTR (sc-8909, Santa Cruz, green fluorescence) in the efferent ductules of Adgrg2 -/Y mice. Scale bars, 50 μm. ( F ) Co-localization of ADGRG2 (red fluorescence) and ezrin (green fluorescence) in the male efferent ductules of WT mice. Scale bars, 50 μm. ( G ) Analysis of ADGRG2 and ezrin fluorescence intensities by Pearson’s correlation analysis. The Pearson's correlation coefficient was 0.69. ( H ) ADGRG2 was immunoprecipitated with an anti-ADGRG2 antibody from the male efferent ductules of WT mice or Adgrg2 -/Y mice, and co-precipitated CFTR, Gs, Gq, β-arrestin-1, β-arrestin-2 and Gi-1/2/3 levels were examined by using specific corresponding antibodies (CFTR antibody:20738–1-AP, Proteintech). (5A-5B) *p < 0.05, **p < 0.01, ***p < 0.001, Adgrg2 -/Y mice compared with WT mice. #p < 0.05, ##p < 0.01, ###p < 0.001. Treatment with selective inhibitors or stimulators was compared with control vehicles. n.s., no significant difference. At least three independent biological replicates were performed for .

Article Snippet: Antibody , Ezrin antibody(rabbit polyclonal) , Proteintech , RRID: AB_2722561 , .

Techniques: Incubation, Quantitative RT-PCR, Fluorescence, Immunofluorescence, Staining, Immunoprecipitation, Control

Journal: eLife

Article Title: Gq activity- and β-arrestin-1 scaffolding-mediated ADGRG2/CFTR coupling are required for male fertility

doi: 10.7554/eLife.33432

Figure Lengend Snippet: ( A ) Diameters of the luminal ductules derived from WT (n = 12), Adgrg2 -/Y (n = 12) or Arrb1 -/- (n = 15) mice. ( B ) Diameters of the luminal ductules derived from WT (n = 12), Adgrg2 -/Y (n = 12) or Arrb2 -/- (n = 15) mice. ( C ) Intracellular pH (pHi) of the ligated efferent ductules derived from WT (n = 9), Arrb1 -/- (n = 9) or Arrb2 -/- (n = 9) mice were measured by carboxy-SNARF. ( D ) Co-localization of ADGRG2 (red fluorescence) and CFTR (sc-8909, Santa Cruz, green fluorescence) in the male efferent ductules of Arrb2 -/- mice. ( E ) Analysis of ADGRG2 and CFTR fluorescence intensities in Arrb2 -/- mice by Pearson’s correlation analysis. The Pearson's correlation coefficient was 0.62. ( F ) Localization of ADGRG2 (red fluorescence) and CFTR (sc-8909, Santa Cruz, green fluorescence) in the male efferent ductules of Arrb1 -/- mice. ( G ) Analysis of ADGRG2 and CFTR fluorescence intensities in Arrb1 -/- mice by Pearson’s correlation analysis. The Pearson's correlation coefficient was −0.15. ( H ) Co-localization of ezrin (red fluorescence) and CFTR (sc-8909, Santa Cruz, green fluorescence) in the male efferent ductules of Arrb2 -/- mice. ( I ) Analysis of ezrin and CFTR fluorescence intensities in Arrb2 -/- mice by Pearson’s correlation analysis. The Pearson's correlation coefficient was 0.66. ( J ) Co-localization of ezrin (red fluorescence) and CFTR (sc-8909, Santa Cruz, green fluorescence) in the male efferent ductules of Arrb1 -/- mice. ( K ) Analysis of ezrin and CFTR fluorescence intensities in Arrb1 -/- mice by Pearson’s correlation analysis. The Pearson's correlation coefficient was −0.15. ( L ) ADGRG2 was immunoprecipitated by an anti-ADGRG2 antibody in the male efferent ductules of Arrb1 -/- mice or Arrb2 -/- mice, and co-precipitates with CFTR, β-arrestin-1, and β-arrestin-2 were examined by using specific corresponding antibodies (CFTR antibody:20738–1-AP, Proteintech). (9A-C) *p < 0.05, **p < 0.01, ***p < 0.001, Adgrg2 -/Y mice compared with WT mice. #p < 0.05, ##p < 0.01, ###p < 0.001, Arrb1 -/- mice or Arrb2 -/- mice compared with WT mice. ns, no significant difference. At least three independent biological replicates were performed for .

Article Snippet: Antibody , Ezrin antibody(rabbit polyclonal) , Proteintech , RRID: AB_2722561 , .

Techniques: Derivative Assay, Fluorescence, Immunoprecipitation

Journal: eLife

Article Title: Gq activity- and β-arrestin-1 scaffolding-mediated ADGRG2/CFTR coupling are required for male fertility

doi: 10.7554/eLife.33432

Figure Lengend Snippet: ( A ) Co-localization of Ezrin (red fluorescence) and CFTR (sc-8909, Santa Cruz, green fluorescence) in male efferent ductules of the WT mice and Adgrg2 -/Y mice. Scale bars, 50 μm. Analysis of Ezrin and CFTR fluorescence intensities by Pearson’s correlation. The pearson's correlation coefficient is 0.6 for WT mice and 0.65 for Adgrg2 -/Y mice. ( B ) Bar graph representation and statistical analyses of . ***p < 0.001, Arrb2 -/- lysates or IP protein were compared with Arrb1 -/- lysates or IP protein, respectively. n.s., no significant difference.

Article Snippet: Antibody , Ezrin antibody(rabbit polyclonal) , Proteintech , RRID: AB_2722561 , .

Techniques: Fluorescence

Journal: eLife

Article Title: Gq activity- and β-arrestin-1 scaffolding-mediated ADGRG2/CFTR coupling are required for male fertility

doi: 10.7554/eLife.33432

Figure Lengend Snippet:

Article Snippet: Antibody , Ezrin antibody(rabbit polyclonal) , Proteintech , RRID: AB_2722561 , .

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Reporter Assay

Journal: Acta biochimica et biophysica Sinica

Article Title: The influence of hyaluronic acid on vascular endothelial cell proliferation and the relationship with ezrin/merlin expression.

doi: 10.1093/abbs/gmr094

Figure Lengend Snippet: Figure 2 The expression and distribution of ezrin and merlin in HUVECs The cells were fixed and visualized by fluorescence microscopy (400). Immunofluorescence stains was shown with red channel. (A) The expression and distribution of ezrin in HUVECs. (B) The expression and distribution of merlin in HUVECs.

Article Snippet: EBM-2 and EGM-2 media were purchased from Lonza (Cambrex, USA); Brdu kits were purchased from Chemicon (Billerica, USA); rabbit anti-human ezrin antibody, rabbit anti-phospho ezrin (Thr567) antibody, rabbit anti-human merlin antibody, and rabbit anti-phospho merlin (Ser518) were obtained from Cell Signaling Technology (Beverly, USA). siRNAs were designed and synthesized by Ambion (Austin, USA).

Techniques: Expressing, Fluorescence, Microscopy, Immunofluorescence

Journal: Acta biochimica et biophysica Sinica

Article Title: The influence of hyaluronic acid on vascular endothelial cell proliferation and the relationship with ezrin/merlin expression.

doi: 10.1093/abbs/gmr094

Figure Lengend Snippet: Figure 3 The effects of oHA and nHA on ezrin/merlin mRNA expression in HUVECs After treatment with 10 mg/ml oHA or 200 mg/ml nHA for different time points, total RNA was extracted and ezrin/merlin mRNA expression was detected by RT–PCR. GAPDH was used a reference. (A, B) The expression of ezrin mRNA after induction by oHA and nHA in HUVECs. (C, D) The expression of merlin mRNA after induction by oHA and nHA in HUVEC. Representative image of three independent experiments was shown. *P , 0.05 vs. PBS group.

Article Snippet: EBM-2 and EGM-2 media were purchased from Lonza (Cambrex, USA); Brdu kits were purchased from Chemicon (Billerica, USA); rabbit anti-human ezrin antibody, rabbit anti-phospho ezrin (Thr567) antibody, rabbit anti-human merlin antibody, and rabbit anti-phospho merlin (Ser518) were obtained from Cell Signaling Technology (Beverly, USA). siRNAs were designed and synthesized by Ambion (Austin, USA).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Acta biochimica et biophysica Sinica

Article Title: The influence of hyaluronic acid on vascular endothelial cell proliferation and the relationship with ezrin/merlin expression.

doi: 10.1093/abbs/gmr094

Figure Lengend Snippet: Figure 4 The effects of oHA and nHA on ezrin/merlin protein expression and their phosphorylation in HUVECs HUVECs were treated with 10 mg/ml oHA or 200 mg/ml nHA for different time points, and total cell lysates were extracted. Protein expression and phosphorylation of ezrin and merlin were examined by western blot using specific antibodies. (A, B, C) The expression of ezrin protein and p-ezrin (Thr567) of HUVECs stimulated by oHA and nHA for different time points. (D, E, F) The expression of merlin protein and p-merlin (Ser518) of HUVECs after induction by oHA and nHA for different time points. Representative image of three independent experiments was shown. GAPDH was used as a reference. *P , 0.05 vs. PBS group.

Article Snippet: EBM-2 and EGM-2 media were purchased from Lonza (Cambrex, USA); Brdu kits were purchased from Chemicon (Billerica, USA); rabbit anti-human ezrin antibody, rabbit anti-phospho ezrin (Thr567) antibody, rabbit anti-human merlin antibody, and rabbit anti-phospho merlin (Ser518) were obtained from Cell Signaling Technology (Beverly, USA). siRNAs were designed and synthesized by Ambion (Austin, USA).

Techniques: Expressing, Phospho-proteomics, Western Blot

Journal: Acta biochimica et biophysica Sinica

Article Title: The influence of hyaluronic acid on vascular endothelial cell proliferation and the relationship with ezrin/merlin expression.

doi: 10.1093/abbs/gmr094

Figure Lengend Snippet: Figure 5 Effects of oHA and nHA on HUVECs proliferation after silencing of ezrin/merlin After HUVECs were transfected with siControl, three siRNA-ezrin or siRNA-merlin sequences for different time points, protein expression of ezrin (A) and merlin (B) was detected by western blot analysis. GAPDH was used as a reference. After silencing of ezrin (C) or merlin (D), HUVECs were stimulated with PBS, 10 mg/ml oHA and 200 mg/ml nHA for 72 h. The number of viable cells was determined using BrDu ELISA method. PBS and siControl are used as controls, respectively. Data were expressed as mean+SD from three different experiments.

Article Snippet: EBM-2 and EGM-2 media were purchased from Lonza (Cambrex, USA); Brdu kits were purchased from Chemicon (Billerica, USA); rabbit anti-human ezrin antibody, rabbit anti-phospho ezrin (Thr567) antibody, rabbit anti-human merlin antibody, and rabbit anti-phospho merlin (Ser518) were obtained from Cell Signaling Technology (Beverly, USA). siRNAs were designed and synthesized by Ambion (Austin, USA).

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

Journal: Acta biochimica et biophysica Sinica

Article Title: The influence of hyaluronic acid on vascular endothelial cell proliferation and the relationship with ezrin/merlin expression.

doi: 10.1093/abbs/gmr094

Figure Lengend Snippet: Figure 6 The effects of oHA and nHA on ezrin/merlin mRNA expression after silencing of ezrin/merlin After ezrin- or merlin-targeting siRNA was transfected into HUVECs, 10 mg/ml oHA or 200 mg/ml nHA was added. After incubation with oHA/nHA for different time points, total RNA was extracted and ezrin/merlin mRNA expression was determined by RT–PCR. (A, B) The effects of oHA and nHA on merlin mRNA expression of HUVECs after transfection of ezrin-targeting siRNA. (C, D) The effects of oHA and nHA on ezrin mRNA expression of HUVECs after transfection of merlin-targeting siRNA. GADPH is used as a reference.

Article Snippet: EBM-2 and EGM-2 media were purchased from Lonza (Cambrex, USA); Brdu kits were purchased from Chemicon (Billerica, USA); rabbit anti-human ezrin antibody, rabbit anti-phospho ezrin (Thr567) antibody, rabbit anti-human merlin antibody, and rabbit anti-phospho merlin (Ser518) were obtained from Cell Signaling Technology (Beverly, USA). siRNAs were designed and synthesized by Ambion (Austin, USA).

Techniques: Expressing, Transfection, Incubation, Reverse Transcription Polymerase Chain Reaction

Journal: Acta biochimica et biophysica Sinica

Article Title: The influence of hyaluronic acid on vascular endothelial cell proliferation and the relationship with ezrin/merlin expression.

doi: 10.1093/abbs/gmr094

Figure Lengend Snippet: Figure 7 The effects of oHA and nHA on ezrin/merlin protein expression and their phosphorylation after silencing of ezrin/merlin After transfection with siRNA-ezrin or siRNA-merlin, HUVECs were treated with 10 mg/ml oHA or 200 mg/ml nHA for 72 h. Then, total cell lysates were extracted and ezrin/merlin protein expression and their phosphorylation levels were detected by western blot analysis. (A, B) The effects of oHA and nHA on merlin protein expression and phosphorylation of HUVECs after transfection with siRNA-ezrin before and 72 h after adding ezrin siRNA. (C, D) The effect of oHA and nHA on ezrin protein expression and phosphorylation of HUVECs after transfection with siRNA-ezrin. GADPH is used as a reference.

Article Snippet: EBM-2 and EGM-2 media were purchased from Lonza (Cambrex, USA); Brdu kits were purchased from Chemicon (Billerica, USA); rabbit anti-human ezrin antibody, rabbit anti-phospho ezrin (Thr567) antibody, rabbit anti-human merlin antibody, and rabbit anti-phospho merlin (Ser518) were obtained from Cell Signaling Technology (Beverly, USA). siRNAs were designed and synthesized by Ambion (Austin, USA).

Techniques: Expressing, Phospho-proteomics, Transfection, Western Blot