Journal: Scientific Data

Article Title: Genome-wide expression datasets of anti-VEGF and dexamethasone treatment of angiogenesis in the rat cornea

doi: 10.1038/sdata.2017.111

Figure Lengend Snippet: (1) sutures were placed intrastromally into the temporal cornea, and immediately followed by topical application of eye drops (IgG, anti- Vegf or dexamethasone). Eye drops were applied until the 48 h time point. At t=48 h, IVCM and slit lamp data was collected and used for phenotypic characterisation. (2) cornea tissue was harvested and used for RNA extraction, and RNA quality verified. (3) high quality RNA was used for target preparation for microarray hybridisation on to GeneChip Rat 230 2.0 microarray chips. The microarray chips were scanned and image files acquired. (4) CEL files were normalised using expression console software. The generated CHP together with the CEL files were submitted to Gene Expression Omnibus repository.

Article Snippet: Three groups of six rats each were treated with one of three topical treatments: IgG (Cat. No. 108-C, R&D Systems) at 20 μgml −1 , anti- Vegf (Cat. No AF 564, a neutralizing rat-specific goat polyclonal pan-VEGFA antibody, R&D Systems, Minneaplois MN, USA) at 20 μgml −1 or dexamethasone (Opnol, Clean Chemical, Sweden AB, Borlänge, Sweden) at 1 mgml −1 .

Techniques: Eye Drops, RNA Extraction, Microarray, Hybridization, Expressing, Software, Generated, Gene Expression

Journal: Scientific Data

Article Title: Genome-wide expression datasets of anti-VEGF and dexamethasone treatment of angiogenesis in the rat cornea

doi: 10.1038/sdata.2017.111

Figure Lengend Snippet: Dataset and sample description across treatment groups.

Article Snippet: Three groups of six rats each were treated with one of three topical treatments: IgG (Cat. No. 108-C, R&D Systems) at 20 μgml −1 , anti- Vegf (Cat. No AF 564, a neutralizing rat-specific goat polyclonal pan-VEGFA antibody, R&D Systems, Minneaplois MN, USA) at 20 μgml −1 or dexamethasone (Opnol, Clean Chemical, Sweden AB, Borlänge, Sweden) at 1 mgml −1 .

Techniques: Microarray, Control

Journal: Scientific Data

Article Title: Genome-wide expression datasets of anti-VEGF and dexamethasone treatment of angiogenesis in the rat cornea

doi: 10.1038/sdata.2017.111

Figure Lengend Snippet: ( a – d ) are signal intensity values correlated between control, IgG, anti- Vegf and dexamethasone treated samples respectively. ( e ) is an example of pathway enrichment analysis and ( f ) is a display of the genes involved in a selected pathway (PI3K-Akt signalling pathway).

Article Snippet: Three groups of six rats each were treated with one of three topical treatments: IgG (Cat. No. 108-C, R&D Systems) at 20 μgml −1 , anti- Vegf (Cat. No AF 564, a neutralizing rat-specific goat polyclonal pan-VEGFA antibody, R&D Systems, Minneaplois MN, USA) at 20 μgml −1 or dexamethasone (Opnol, Clean Chemical, Sweden AB, Borlänge, Sweden) at 1 mgml −1 .

Techniques: Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 was upregulated in oral squamous cell carcinoma (OSCC) tissue and associated with clinical disease stages in human OSCC. (A) Correlation CX3CL1 gene expression with the normal and tumour cells using GEO microarray 3524 OSCC tissue samples. (B) CX3CL1 expression profiles in 497 OSCC tissue specimens were analysed obtained from the Cancer Genome Atlas (TCGA) database. (C,D) OSCC specimens were subjected to IHC staining. (E) Kaplan–Meier survival analysis of the associations between high or low plasma levels of CX3CL1 expression and overall survival of OSCC patients. * p < 0.05 compared with controls.

Article Snippet: Specific neutralizing antibodies for CX3CL1 (AF365‐SP) and CX3CR1 (AF5825) were acquired from Novus Biologicals.

Techniques: Gene Expression, Microarray, Expressing, Immunohistochemistry, Clinical Proteomics

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 upregulates human oral squamous cell carcinoma (OSCC) cell migration and invasion. (A‐C) OSCC cells were incubated with different concentrations of CX3CL1 for 24 h; then, cell migration was assessed using the (A) in vitro wound‐healing assay, (B,C) the Transwell assay. (D) Quantified result of cell migration with CX3CL1 neutralizing antibody. (E) Quantified result of cell invasion with CX3CL1 neutralizing antibody. Results are expressed as the mean ± SD of four independent experiments. * p < 0.05 as compared with controls.

Article Snippet: Specific neutralizing antibodies for CX3CL1 (AF365‐SP) and CX3CR1 (AF5825) were acquired from Novus Biologicals.

Techniques: Migration, Incubation, In Vitro, Wound Healing Assay, Transwell Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 activates tumour cell migration via the ICAM‐1 expression in human oral squamous cell carcinoma (OSCC) cells. (A) ICAM‐1 expression in the normal and tumour cells obtained from the TCGA dataset analysis. (B) VCAM‐1 expression in the normal and tumour cells obtained from the TCGA dataset analysis. (C) Correlation analysis of CX3CL1 and ICAM‐1 expression using the TIMER2.0 database. (D) Correlation analysis of CX3CL1 and VCAM‐1 expression using the TIMER2.0 database. (E) Quantified result of ICAM‐1 gene expression with CX3CL1 recombinant protein (30 ng/mL) treatment. (F) Quantified result of VCAM‐1 gene expression with CX3CL1 recombinant protein (30 ng/mL) treatment. (G) Protein expression of ICAM‐1 with concentration‐depended CX3CL1 treatment. (H) Protein expression of VCAM‐1 with concentration‐depended CX3CL1 treatment. (I) Quantified result of cell migration with ICAM‐1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. (J) Quantified result of cell migration with ICAM‐1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Specific neutralizing antibodies for CX3CL1 (AF365‐SP) and CX3CR1 (AF5825) were acquired from Novus Biologicals.

Techniques: Migration, Expressing, Gene Expression, Recombinant, Concentration Assay, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 upregulates cell motility and ICAM‐1 expression via its receptor CX3CR1. (A) Levels of CX3CR1 mRNA expression in normal tongue tissue and human oral squamous cell carcinoma (OSCC) tumour tissue were analysed using records from the GEO data set GSE13601. (B) Levels of CX3CR1 mRNA expression in different N stages of OSCC tumour tissue were analysed using records from the GEO data set GSE78060. (C) Quantified result of cell movement with CX3CR1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. (D) Quantified result of cell migration with CX3CR1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. (E) Quantified resulting of ICAM‐1 expression with CX3CR1 neutralizing antibody and CX3CL1 recombinant protein (30 ng/mL) treatment. (F) Quantified result of cell movement with CX3CR1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. (G) Quantified result of cell migration with CX3CR1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. (H) Quantified result of ICAM‐1 expression with CX3CR1 siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Specific neutralizing antibodies for CX3CL1 (AF365‐SP) and CX3CR1 (AF5825) were acquired from Novus Biologicals.

Techniques: Expressing, Recombinant, Migration, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 triggers the phosphorylation of PLCβ/PKCα/c‐Src to signal the cell movement, migration and ICAM‐1 expression in human oral squamous cell carcinoma (OSCC) cells. (A‐C) Quantified result of cell movement, cell migration and ICAM‐1 expression with signalling inhibitors and CX3CL1 recombinant protein (30 ng/mL) treatment. (D‐E) Protein expression of phosphorylated PLCβ, PKCα and c‐Src with CX3CL1 recombinant protein (30 ng/mL) treatment in SCC4 cell and SAS cell. (F‐H) Quantified result of cell movement, cell migration and ICAM‐1 expression with PLCβ, PKCα or c‐Src siRNA and CX3CL1 recombinant protein (30 ng/mL) treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Specific neutralizing antibodies for CX3CL1 (AF365‐SP) and CX3CR1 (AF5825) were acquired from Novus Biologicals.

Techniques: Phospho-proteomics, Migration, Expressing, Recombinant, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 promotes the phosphorylation of c‐Jun to upregulate the cell motility and ICAM‐1 expression in human oral squamous cell carcinoma (OSCC) cells. (A) Quantified result of CX3CL1‐induced cell movement with c‐Jun inhibitors (B) Quantified result of CX3CL1‐induced cell migration with c‐Jun inhibitors (C) Quantified result of CX3CL1‐induced ICAM‐1 expression with c‐Jun inhibitors (D) Protein expression of phosphorylated c‐Jun with CX3CL1 recombinant protein (30 ng/mL) treatment. (E) Quantified result of CX3CL1‐induced cell movement with c‐Jun siRNA treatment. (F) Quantified result of CX3CL1‐induced cell migration with c‐Jun siRNA treatment. (G) Quantified result of CX3CL1‐induced ICAM‐1 expression with c‐Jun siRNA treatment. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Specific neutralizing antibodies for CX3CL1 (AF365‐SP) and CX3CR1 (AF5825) were acquired from Novus Biologicals.

Techniques: Phospho-proteomics, Expressing, Migration, Recombinant, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: CX3CL1 induces cell migration and invasion through ICAM ‐1 expression in oral squamous cell carcinoma cells

doi: 10.1111/jcmm.17750

Figure Lengend Snippet: CX3CL1 promotes the translocation of c‐Jun and promoter binding of AP‐1 formation in human oral squamous cell carcinoma (OSCC) cells. (A) Immunofluorescent staining showed that c‐Jun translocated into the nucleus after cell were treated with CX3CL1; nuclear translocation was prevented when the cells were pre‐treated with CX3CR1 mAb, U73122, GF109203 or PP2. (B) Quantified analysis of c‐Jun activation with CX3CL1 using Western blot. (C) CX3CL1 treatment upregulated AP‐1 luciferase activity. (D) Quantified analysis of AP‐1 luciferase activity with specific inhibitors and CX3CL1 recombinant protein (30 ng/mL) treatment. (E) Chromatin immunoprecipitation analysis of c‐Jun and AP‐1 promoter binding site with neutralizing antibody and specific inhibitors. Results are expressed as the mean ± SD of four independent experiments. *, p < 0.05 and #, p < 0.05 as compared to control and CX3CL1 treatment.

Article Snippet: Specific neutralizing antibodies for CX3CL1 (AF365‐SP) and CX3CR1 (AF5825) were acquired from Novus Biologicals.

Techniques: Translocation Assay, Binding Assay, Staining, Activation Assay, Western Blot, Luciferase, Activity Assay, Recombinant, Chromatin Immunoprecipitation, Control

Journal: Experimental Biology and Medicine

Article Title: Annona atemoya leaf extract ameliorates cognitive impairment in amyloid-β injected Alzheimer’s disease-like mouse model

doi: 10.1177/1535370219886269

Figure Lengend Snippet: Molecular mechanisms responsible for anti-AD effects of AAL extract. (a) HT22 cells were exposed to H2O2 in the absence or presence of AAL extract (50 μg/mL) for 6 h. Antibody microarray assay was performed using the phospho-specific antibody microarray slide (Full Moon BioSystems). For data acquisition, GenePix 4100A scanner (Axon Instrument, USA) was used. The normalization data were analyzed using Genowiz 4.0™ (Ocimum Biosolutions). The phosphorylation ratio was calculated and represented as fold changes of indicated phosphoproteins after H2O2 treatment normalized to total protein expression (upper panel). Total protein quantification is shown (lower panel). (b) HT22 cells were exposed to H2O2 with or without various concentrations of AAL extract (0, 12.5, 25, or 50 μg/mL) for 6 h. Cell lysates were prepared from HT22 cells and equal amounts of protein were subjected to Western blotting using anti-phospho-CaMK2 β/ν/δ, GRK2, EGFR, Myc, FER, caveolin-1, NFκB p65, and MLRN 2 antibodies to validate the Ab microarray. (c) Protein extracts were prepared from hippocampal tissues in an Aβ-induced AD mouse model. Vehicle or various concentrations of AAL extract (50, 100, or 200 mg/kg) were administered to Aβ mice for 23 days. Western blotting was performed for anti-phospho-EGFR and phospho-GRK2. The validity of the two phospho-antibodies was determined using pre-stained protein marker (Bio-Rad). GAPDH was used as an internal control. Shown blots are representative results from three independent experiments.

Article Snippet: Molecular mechanisms responsible for anti-AD effects of AAL extract. (a) HT22 cells were exposed to H 2 O 2 in the absence or presence of AAL extract (50 μg/mL) for 6 h. Antibody microarray assay was performed using the phospho-specific antibody microarray slide (Full Moon BioSystems).

Techniques: Microarray, Phospho-proteomics, Expressing, Western Blot, Staining, Marker, Control

Journal: Cell communication and signaling : CCS

Article Title: Epithelial and interstitial Notch1 activity contributes to the myofibroblastic phenotype and fibrosis.

doi: 10.1186/s12964-019-0455-y

Figure Lengend Snippet: Fig. 1 Epithelial and interstitial Notch1 activity is associated with the myofibroblastic phenotype in UUO kidneys. a Enhanced expression of Ki67 in UUO kidneys determined by immunohistochemical staining. Bar = 50 μm. b Upregulated Ki67 expression occurred in the tubules and interstitium of the kidney. c Increased expression of Histone H3 (phosphor S10) and c-Myc in UUO kidneys determined by western blot. d The mRNA expression levels of c-Myc and P53 were augmented in UUO kidneys determined by qRT-PCR. e Immunohistochemical staining showed that the expression levels of Notch1 and NICD were elevated in UUO kidneys. Bar = 50 μm. f NICD expression was upregulated in the tubules and interstitium of kidneys. g mRNA microarray revealed enhanced expression of Notch1-associated genes in UUO kidneys. h The expression levels of NICD, Notch1 and Jagged1 in UUO kidneys were increased. i Increased mRNA expression levels of Hes1, Hes6, Hey1, Hey2, and Numb in UUO kidneys were observed. j Elevated expression of Notch1, NICD and α-SMA determined by immunofluorescence staining in renal tubules and interstitium of CKD patients. All data are presented as means ± SDs. *P < 0.05, **P < 0.01 versus the sham group

Article Snippet: Immunofluorescence staining was performed using anti-Ki67, α-SMA, NICD, Notch1, Jagged1, and collagen III as the primary antibodies, and the cell preparations were incubated with Alexa Fluor 488 (green)- or 594 (red) (Proteintech)-labelled secondary antibodies.

Techniques: Activity Assay, Expressing, Immunohistochemical staining, Staining, Western Blot, Quantitative RT-PCR, Microarray, Immunofluorescence

Journal: Cell communication and signaling : CCS

Article Title: Epithelial and interstitial Notch1 activity contributes to the myofibroblastic phenotype and fibrosis.

doi: 10.1186/s12964-019-0455-y

Figure Lengend Snippet: Fig. 2 Notch1 signalling is activated in TECs after AA injury. a RTCA assay indicating the survival index of NRK-52E cells treated with AA with or without DAPT. b Upregulated Ki67 expression, determined by immunofluorescence staining, was inhibited by DAPT treatment. Bar = 20 μm. c The ratio of Ki67-positive cells in total cells according to the results of immunofluorescence staining. d qRT-PCR showed that increased mRNA expression levels of Col1α1, Col3α1 and α-SMA, and decreased mRNA levels of E-cadherin and BMP-7 in AA-treated NRK-52E cells were inhibited by DAPT treatment. e Elevated expression levels of α-SMA and collagen III in AA-treated NRK-52E cells were inhibited by DAPT treatment. Bar = 20 μm. f Increased expression levels of α-SMA, vimentin and collagen I, and decreased expression of E-cadherin in AA-treated NRK-52E cells were inhibited by DAPT treatment. g Increased mRNA expression levels of Notch1, Jagged1 and Numb in AA-treated NRK-52E cells were inhibited by DAPT treatment. h Elevated expression levels of Notch1, NICD and Jagged1 in AA-treated NRK-52E cells were inhibited by DAPT treatment. Bar = 20 μm. i Enhanced expression levels of Notch1, Jagged1 and NICD in AA-treated NRK-52E cells were inhibited by DAPT treatment. All data are presented as means ± SDs. AA, 10 ng/ml; DAPT (HD), high-dose (10 μmol/L); DAPT (LD), low-dose (1 μmol/L). *P < 0.05, **P < 0.01 versus the control; #P < 0.01, #P < 0.05 versus the AA-treated group

Article Snippet: Immunofluorescence staining was performed using anti-Ki67, α-SMA, NICD, Notch1, Jagged1, and collagen III as the primary antibodies, and the cell preparations were incubated with Alexa Fluor 488 (green)- or 594 (red) (Proteintech)-labelled secondary antibodies.

Techniques: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Control

Journal: Cell communication and signaling : CCS

Article Title: Epithelial and interstitial Notch1 activity contributes to the myofibroblastic phenotype and fibrosis.

doi: 10.1186/s12964-019-0455-y

Figure Lengend Snippet: Fig. 3 Notch1 signalling is activated in fibroblasts after AA injury. a RTCA assay indicated the survival index of NRK-49F cells treated with AA with or without DAPT. b Upregulated Ki67 expression, determined by immunofluorescence staining, was inhibited by DAPT treatment. Bar = 20 μm. c Ratio of Ki67-positive cells in total cells according to the results of immunofluorescence staining. d qRT-PCR showed that increased mRNA expression levels of Col3α1 and α-SMA in AA-treated NRK-49F cells were inhibited by DAPT treatment. e Increased expression levels of α-SMA, vimentin and collagen I in AA-treated NRK-49F cells were inhibited by DAPT treatment. f Elevated expression levels of α-SMA and collagen III in AA-treated NRK-49F cells were inhibited by DAPT treatment. Bar = 20 μm. g Increased mRNA expression levels of Notch1 and Numb in AA-treated NRK-49F cells were inhibited by DAPT treatment. h Elevated expression of NICD in AA-treated NRK-49F cells was inhibited by DAPT treatment. Bar = 20 μm. All data are presented as means ± SDs. AA, 10 ng/ml; DAPT (HD), high-dose (10 μmol/L); DAPT (LD), low-dose (1 μmol/L). *P < 0.05 versus the control; #P < 0.01 versus the AA-treated group

Article Snippet: Immunofluorescence staining was performed using anti-Ki67, α-SMA, NICD, Notch1, Jagged1, and collagen III as the primary antibodies, and the cell preparations were incubated with Alexa Fluor 488 (green)- or 594 (red) (Proteintech)-labelled secondary antibodies.

Techniques: Expressing, Immunofluorescence, Staining, Quantitative RT-PCR, Control

Journal: Cell communication and signaling : CCS

Article Title: Epithelial and interstitial Notch1 activity contributes to the myofibroblastic phenotype and fibrosis.

doi: 10.1186/s12964-019-0455-y

Figure Lengend Snippet: Fig. 4 Activated Notch1 signalling induces myofibroblastic phenotype in vitro. a Increased mRNA expression levels of Notch1 and Numb in Jag1- fc-treated NRK-52E and NRK-49F cells were strengthened by TGF-β1. b Upregulated NICD expression and nuclear localization in Jag1-fc-treated NRK-52E and NRK-49F cells were strengthened by TGF-β1. Bar = 20 μm. c Increased mRNA expression levels of Col3α1 and α-SMA in Jag1-fc- treated NRK-52E and NRK-49F cells were enhanced by TGF-β1. d Increased expression of α-SMA and vimentin in Jag1-fc-treated NRK-52E cells or NRK-49F cells were enhanced by TGF-β1. e Elevated expression of α-SMA and collagen III in Jag1-fc-treated NRK-52E and NRK-49F cells was strengthened by TGF-β1. Bar = 20 μm. All data are presented as means ± SDs. Jag1-fc, 1 μg/ml; TGF-β1, 5 ng/ml. *P < 0.05, **P < 0.05 versus the control; #P < 0.05 versus the Jag1-fc-treated group

Article Snippet: Immunofluorescence staining was performed using anti-Ki67, α-SMA, NICD, Notch1, Jagged1, and collagen III as the primary antibodies, and the cell preparations were incubated with Alexa Fluor 488 (green)- or 594 (red) (Proteintech)-labelled secondary antibodies.

Techniques: In Vitro, Expressing, Control

Journal: Cell communication and signaling : CCS

Article Title: Epithelial and interstitial Notch1 activity contributes to the myofibroblastic phenotype and fibrosis.

doi: 10.1186/s12964-019-0455-y

Figure Lengend Snippet: Fig. 6 TGF-β1 induces Notch1 signalling activation during the EMT and FMD in vitro. a RTCA assay indicated the survival index of NRK-52E cells treated with TGF-β1 with or without DAPT. b RTCA assay indicated the survival index of NRK-49F cells treated with TGF-β1 with or without DAPT. c Increased mRNA expression levels of Notch1, Jagged1, Hey1, Col1α1, Col3α1, and α-SMA in TGF-β1-treated NRK-52E cells were inhibited by DAPT. d Increased mRNA expression levels of Notch1, Jagged1, Hey1, Col1α1, Col3α1, and α-SMA in TGF-β1-treated NRK-49F cells were inhibited by DAPT. e Elevated expression levels of NICD, Notch1 and Jagged1 in TGF-β1-treated NRK-52E cells were inhibited by DAPT. f Elevated expression levels of NICD, Notch1 and Jagged1 in TGF-β1-treated NRK-49F cells were inhibited by DAPT. All data are presented as means ± SDs. TGF-β1, 5 ng/ml. DAPT (HD), high-dose (10 μmol/L); DAPT (LD), low-dose (1 μmol/L). *P < 0.05, versus the control; #P < 0.05 versus the TGF-β1-treated group

Article Snippet: Immunofluorescence staining was performed using anti-Ki67, α-SMA, NICD, Notch1, Jagged1, and collagen III as the primary antibodies, and the cell preparations were incubated with Alexa Fluor 488 (green)- or 594 (red) (Proteintech)-labelled secondary antibodies.

Techniques: Activation Assay, In Vitro, Expressing, Control

Journal: Cell communication and signaling : CCS

Article Title: Epithelial and interstitial Notch1 activity contributes to the myofibroblastic phenotype and fibrosis.

doi: 10.1186/s12964-019-0455-y

Figure Lengend Snippet: Fig. 7 Notch1 activity mediates myofibroblastic phenotype dependent on TGF-β1 in vitro. a The mRNA expression levels of TGF-β1R, Col3α1 and α-SMA in NRK52E and NRK-49F cells treated with TGF-β1 with or without pcDNA3.1-NICD plasmid or NICD siRNA. b The protein expression levels of TGF-β1R, α-SMA and collagen III in NRK52E and NRK-49F cells treated with TGF-β1 with or without pcDNA3.1-NICD plasmid or NICD siRNA. c The mRNA expression levels of Smad2 and Smad3 in NRK52E and NRK-49F cells treated with TGF-β1 with or without pcDNA3.1-NICD plasmid or NICD siRNA. d The protein expression and phosphorylation levels of Smad2 and Smad3 in NRK52E and NRK-49F cells treated with TGF-β1 with or without pcDNA3.1-NICD plasmid or NICD siRNA. All data are presented as means ± SDs. TGF-β1, 5 ng/ml. *P < 0.05, versus the control; #P < 0.05 versus the TGF-β1-treated group; §P < 0.05 versus the pcDNA3.1-NICD plasmid-treated group

Article Snippet: Immunofluorescence staining was performed using anti-Ki67, α-SMA, NICD, Notch1, Jagged1, and collagen III as the primary antibodies, and the cell preparations were incubated with Alexa Fluor 488 (green)- or 594 (red) (Proteintech)-labelled secondary antibodies.

Techniques: Activity Assay, In Vitro, Expressing, Plasmid Preparation, Phospho-proteomics, Control

Journal: Bioactive Materials

Article Title: Bioinspired engineering ADSC nanovesicles thermosensitive hydrogel enhance autophagy of dermal papilla cells for androgenetic alopecia treatment

doi: 10.1016/j.bioactmat.2024.02.023

Figure Lengend Snippet: JAM-A protein plays a core role in ADSC-NVs-mediated AGA hair regrowth. A) Volcano plot of the GSE184501 microarray data set, and JAM-A significantly downgraded. B) Change in JAM-A expression. C) Relative expression of JAM-A. D) Viability of DPC cell lines treated with DHT for 48 h. E) Statistical data on the proportion of senescent cells in each group was counted in the same random area. F) Effect of DHT on DPC cell line migration at every indicating time; scale bar: 50 μm. G) Relative migration of DHT-injured DPC cell lines in each group. H) Apoptosis cells of DPC cell lines were injured with macrophages for 48 h. I) Statistical data on the proportion of apoptosis cells in each group. J) Change in LC3, AKT, AKT phosphorylation, and p62 expression. K) Relative expression of LC-3II, p62, and AKT phosphorylation. L) Change in Cyclin D1 and β-catenin expression. M) Relative expression of Cyclin D1 and β-catenin. (n = 3 per group, *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: Subsequently, they were incubated with CD63 (1:1000, ab134045, Abcam) or TSG101 (1:1000, ab125011, Abcam) or Calnexin (1:1000, ab133615, Abcam) or β-catenin (1:1000, ab32572, Abcam) or LC3B (1:1000, ab192890, Abcam) or GAPDH (1:4000, 60,004-1-lg, Proteintech) or α-tubulin (1:1000, 2144S, Cell Signaling Technology) or JAM-A (1:1000, 36–1700, Invitrogen) or p62 (1:1000, ab109012, Abcam) or β-actin (1:1000, GB11001-100, Servicebio) or AKT (1:1000, 4685, Cell Signaling Technology) or phospho-Akt (1:1000, 4060S, Cell Signaling Technology) or Cyclin D1 (1:1000, 60186-1-Ig, Proteintech) primary antibodies overnight at 4 °C.

Techniques: Microarray, Expressing, Migration, Phospho-proteomics

Journal: Bioactive Materials

Article Title: Bioinspired engineering ADSC nanovesicles thermosensitive hydrogel enhance autophagy of dermal papilla cells for androgenetic alopecia treatment

doi: 10.1016/j.bioactmat.2024.02.023

Figure Lengend Snippet: Protective effect of JAM-A OE @NV on JAM-A-downregulated DPC cell line injured by DHT and macrophages. A) Change in JAM-A expression. B) Relative expression of JAM-A. C) Viability of the DHT-injured, downregulated DPC cell line treatment with JAM-A@NV for 48 h. D) Statistical data on the proportion of senescent cells in each group was counted in the same random area. E) Effect of JAM-A@NV on the DHT-injured, downregulated DPC cell line migration at 24 h; scale bar: 50 μm. F) Relative migration of the DHT-injured, downregulated DPC cell line in each group. G) Apoptosis cells of the macrophages injured by downregulated DPC cell line treatment with JAM-A@NV for 48 h. H) Statistical data on the proportion of apoptosis cells in each group. I) Change in LC3, AKT, AKT phosphorylation, and p62 expression. J) Relative expression of LC-3II, p62, and AKT phosphorylation. K) Change in Cyclin D1 and β-catenin expression. L) Relative expression of Cyclin D1 and β-catenin. (n = 6 per group, *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: Subsequently, they were incubated with CD63 (1:1000, ab134045, Abcam) or TSG101 (1:1000, ab125011, Abcam) or Calnexin (1:1000, ab133615, Abcam) or β-catenin (1:1000, ab32572, Abcam) or LC3B (1:1000, ab192890, Abcam) or GAPDH (1:4000, 60,004-1-lg, Proteintech) or α-tubulin (1:1000, 2144S, Cell Signaling Technology) or JAM-A (1:1000, 36–1700, Invitrogen) or p62 (1:1000, ab109012, Abcam) or β-actin (1:1000, GB11001-100, Servicebio) or AKT (1:1000, 4685, Cell Signaling Technology) or phospho-Akt (1:1000, 4060S, Cell Signaling Technology) or Cyclin D1 (1:1000, 60186-1-Ig, Proteintech) primary antibodies overnight at 4 °C.

Techniques: Expressing, Migration, Phospho-proteomics

Journal: Blood

Article Title: Jun-regulated genes promote interaction of diffuse large B-cell lymphoma with the microenvironment

doi: 10.1182/blood-2014-04-568188

Figure Lengend Snippet: Immunohistochemical analysis of p-c-Jun (Ser73) expression in a subset of 25 DLBCL tumor tissues collected from patients with or without bone marrow involvement

Article Snippet: Antibody specific for p-c-Jun (Ser73) was purchased from Cell Signaling (#9164).

Techniques: Immunohistochemical staining, Expressing, Staining

Journal: Blood

Article Title: Jun-regulated genes promote interaction of diffuse large B-cell lymphoma with the microenvironment

doi: 10.1182/blood-2014-04-568188

Figure Lengend Snippet: c-Jun is enriched in extranodal lymphoma. (A) Positron emission tomography/computed tomography (PET/CT) images from patients with different lymphoma localization. Patients #1 and #2 had only nodal sites of involvement; representative images are shown. Patient #3 had multifocal extranodal involvement in the bone marrow in the spine, as shown in sagittal images. Patient #4 had multiple extranodal sites of involvement, including bone marrow, subcutaneous tissues, and perineural spread. Expression of p-c-Jun (Ser73) in DLBCL tumor tissues from these patients was determined by immunohistochemistry; representative images are shown (×500). (B-D) Expression of c-Jun was stratified by the number of extranodal sites using microarray data available in a public repository (GSE10846). Log2 median-centered intensities of c-Jun (probe 201465_s) in 296 cases with a known number of extranodal sites were obtained by using Oncomine v4.5 (www.oncomine.org). Patients diagnosed with the ABC or GCB subtypes of DLBCL were analyzed in (C) and (D), respectively. Statistical significance was evaluated by using two-tailed Student t test. ****P < .0001; ***P < .001; **P < .01; *P < .05.

Article Snippet: Antibody specific for p-c-Jun (Ser73) was purchased from Cell Signaling (#9164).

Techniques: Positron Emission Tomography, Computed Tomography, Positron Emission Tomography-Computed Tomography, Expressing, Immunohistochemistry, Microarray, Two Tailed Test

Journal: Immunity

Article Title: T-bet transcription factor promotes antibody secreting cell differentiation by limiting the inflammatory effects of IFNγ on B cells

doi: 10.1016/j.immuni.2019.04.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Bound Ab was detected with AP-conjugated goat anti-mouse heavy chain-specific pan-IgG Ab (Jackson ImmunoResearch) and the AP substrate BCIP/NBT (Moss Substrates).

Techniques: Blocking Assay, Recombinant, Purification, Expressing, Staining, Labeling, Microarray, Software

Journal: Molecular Cancer Therapeutics

Article Title: Suppression of Extracellular Vesicle VEGF-C–mediated Lymphangiogenesis and Pancreatic Cancer Early Dissemination By a Selective HDAC1/2 Inhibitor

doi: 10.1158/1535-7163.mct-20-0963

Figure Lengend Snippet: Figure 2. Exogenous expression of DUSP2 inhibits pancreatic tumor formation. A, Comparative transcriptome profiling of HDAC inhibitors. Microarray analysis of upregulated gene expression in TSA (T) and sodium butyrate (B) treated cells. Two time-points (12 and 48 hours) were included for analysis. B, RT-qPCR of DUSP2 expression in MIA PaCa-2 cells treated with SAHA (S, 4 mmol/L) and B390 (B, 0.25 and 1 mmol/L) for 24 hours. CYCLOPHILIN B (PPIB) was used as internal control. C, Western blotting result for the expression of DUSP2, ERK1/2 phosphorylation (pERK) and expression (tERK) in MIA PaCa-2 cells treated with control or B390 (0.25 and 1 mmol/L) for 24 hours. GAPDH were detected as internal control for whole cell lysate. D, Representative images show the fluorescent signal of GFP in GFP or DUSP2- GFP–transfected MIA PaCa-2 cells (left). Western blotting for the expression of DUSP2 and ERK1/2 in MIA PaCa-2 cells transfected with GFP or DUSP2-GFP expression vector. GAPDH is the internal control for whole-cell lysate (right). E, Representative images show crystal violet-stained cells growing on 24-well plates for 48 hours. Equal numbers of MIA PaCa-2 cells were plated on 24-well plates and transfected with GFP or DUSP2-GFP expression vectors. This experiment was repeated by MTT assay with similar result (right). F, Western blotting for the expression of cleaved caspase 3 (cCas3) in MIA PaCa-2 cells. MIA PaCa-2 cells were transfected with GFP or DUSP2-GFP and treated with DMSO (D) or B390 (1 mmol/L) for 24 hours. b-Actin is the loading control. G, Growth curve (left), tumor mass, and representative image (right) show forced expression of DUSP2 abolished tumor growth in mice pancreas. GFP or DUSP2-GFP MIA PaCa-2 cells were labeled with luciferase and injected into mouse pancreas (n ¼ 6 mice per group). Tumor masswas followed by IVIS imaging. 42 days after inoculation, mice were sacrificed, tumors and pancreases were collected. H, IHC of Ki67 and cleaved caspase 3 in control (shCtrl) and DUSP2 KD (shDUSP2) PANC-1 tumors. , P < 0.05; , P < 0.001.

Article Snippet: Antibodies against DUSP2 (Santa Cruz Biotechnology, catalog no. sc-32776, RRID:AB_2094883), HDAC1 (Cell Signaling Technology, catalog no. 5356, RRID: AB_10612242), HDAC2 (Cell Signaling Technology, catalog no. 5113, RRID:AB_10624871), phospho-p44/42 MAPK (Cell Signaling Technology, catalog no. 4370, RRID:AB_2315112), p44/42MAPK (Cell Signaling Technology, catalog no. 4696, RRID:AB_390780), VEGF-C (GeneTex, GTX113574, RRID:AB_10620764), CD9 (ProteinTech, 60232–1, RRID: AB_11232215),HSP70 (SantaCruzBiotechnology, catalogno. sc-32239), GAPDH (GeneTex, GTX100118, RRID:AB_1080976) were used.

Techniques: Expressing, Microarray, Gene Expression, Quantitative RT-PCR, Control, Western Blot, Phospho-proteomics, Transfection, Plasmid Preparation, Staining, MTT Assay, Labeling, Luciferase, Injection, Imaging

Journal: Molecular Cancer Therapeutics

Article Title: Suppression of Extracellular Vesicle VEGF-C–mediated Lymphangiogenesis and Pancreatic Cancer Early Dissemination By a Selective HDAC1/2 Inhibitor

doi: 10.1158/1535-7163.mct-20-0963

Figure Lengend Snippet: Figure 3. Loss of Dusp2 in the pancreas facilitates the progression of Kras-driven PanIN progression. A, Schematic diagram of breeding KC and KDC transgenic mouse model. B, Histology of pancreas from 2-month old KDC mice and WT littermate. C, H&E staining of pancreas in 6-month old WT (a and b), KC mice (c and d), and KDC mice (e and f). Normal pancreatic ducts (b) show single layer of epithelial cells. Pancreas of KC shows ADM and PanINs while pancreas of KDC display larger area of abnormalities, including ADM, PanINs, and carcinoma. Early PanIN display papillary or micropapillary projections. Advanced PanIN has epithelial cells budding into the lumen and necrosis in the lumen. Carcinoma displays nuclear abnormalities and glands embedded in tumor stroma. D, IHC staining of phosphor- p44/42 MAPK (pERK) in 6-month old pancreas from control, KC, and KDC mice.

Article Snippet: Antibodies against DUSP2 (Santa Cruz Biotechnology, catalog no. sc-32776, RRID:AB_2094883), HDAC1 (Cell Signaling Technology, catalog no. 5356, RRID: AB_10612242), HDAC2 (Cell Signaling Technology, catalog no. 5113, RRID:AB_10624871), phospho-p44/42 MAPK (Cell Signaling Technology, catalog no. 4370, RRID:AB_2315112), p44/42MAPK (Cell Signaling Technology, catalog no. 4696, RRID:AB_390780), VEGF-C (GeneTex, GTX113574, RRID:AB_10620764), CD9 (ProteinTech, 60232–1, RRID: AB_11232215),HSP70 (SantaCruzBiotechnology, catalogno. sc-32239), GAPDH (GeneTex, GTX100118, RRID:AB_1080976) were used.

Techniques: Transgenic Assay, Staining, Immunohistochemistry, Control

Journal: The Journal of Biological Chemistry

Article Title: Noncanonical Transforming Growth Factor β (TGFβ) Signaling in Cranial Neural Crest Cells Causes Tongue Muscle Developmental Defects * ^♦

doi: 10.1074/jbc.M113.493551

Figure Lengend Snippet: Altered TGFβ signaling pathway in the tongue of Tgfbr2fl/fl;Wnt1-Cre mice. A, the table lists phosphorylated proteins altered in Tgfbr2fl/fl;Wnt1-Cre tongue as compared with controls expressed as a ratio of phosphorylated protein over nonphosphorylated protein (P/N). Ab, antibody. B, immunoblotting analysis of ABL1 and PKC and their phosphorylated forms (P) in the tongues of E14.5 control (WT; lane 1) and Tgfbr2fl/fl;Wnt1-Cre (CKO; lane 2) mice. C, immunoblotting analysis of the indicated molecules in primary mouse tongue mesenchymal cells from E14.5 control (WT) and Tgfbr2fl/fl;Wnt1-Cre (CKO) mice after treatment with TGFβ1 for 0, 10, 30, and 60 min.

Article Snippet: Phosphoprotein Profiling by the TGFβ Signaling Phospho-specific Antibody Microarray The TGFβ signaling phospho-specific antibody microarray, which was designed and manufactured by Full Moon Biosystems, Inc. (Sunnyvale, CA), contains 176 highly specific and well characterized phosphorylation antibodies.

Techniques: Western Blot, Control

Journal: The Journal of Biological Chemistry

Article Title: Noncanonical Transforming Growth Factor β (TGFβ) Signaling in Cranial Neural Crest Cells Causes Tongue Muscle Developmental Defects * ^♦

doi: 10.1074/jbc.M113.493551

Figure Lengend Snippet: Ectopic activation of noncanonical TGFβ signaling is prevented by reduction of TβRI/ALK5 in Tgfbr2fl/fl;Wnt1-Cre mice. A and B, immunoblotting analysis of ABL1, phosphorylated (P) ABL1, and tenascin C in tongues from E14.5 control (lane 1), Tgfbr2fl/fl;Wnt1-Cre (lane 2), and Tgfbr2fl/fl;Wnt1-Cre;Alk5fl/+ (lane 3) mice. C, quantitative RT-PCR analyses of tenascin C (TnC) in E14.5 tongues of Tgfbr2fl/fl control (blue bars), Tgfbr2fl/fl;Wnt1-Cre (red bars), and Tgfbr2fl/fl;Wnt1-Cre;Alk5fl/+ (green bars) mice. ***, p < 0.001. D, Gomori's aldehyde fuchsin staining in the tongue of Tgfbr2fl/fl;Wnt1-Cre;Alk5fl/+ mice at E18.5. Purple, elastic fibers; green, collagen fibers and nuclei; red, muscle. Scale bar, 500 μm. E, AZAN staining in the tongue of Tgfbr2fl/fl;Wnt1-Cre;Alk5fl/+ mice at E18.5. Blue, collagen fibers; red, muscle; orange, nuclei. Bar, 500 μm. F, quantitative RT-PCR analyses of the indicated molecules in E14.5 tongues of Tgfbr2fl/fl control (blue bars), Tgfbr2fl/fl;Wnt1-Cre (red bars), and Tgfbr2fl/fl;Wnt1-Cre;Alk5fl/+ (green bars) mice. ***, p < 0.001, *, p < 0.05.

Article Snippet: Phosphoprotein Profiling by the TGFβ Signaling Phospho-specific Antibody Microarray The TGFβ signaling phospho-specific antibody microarray, which was designed and manufactured by Full Moon Biosystems, Inc. (Sunnyvale, CA), contains 176 highly specific and well characterized phosphorylation antibodies.

Techniques: Activation Assay, Western Blot, Control, Quantitative RT-PCR, Staining