Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 1. Increased expression of PEDF in expanded human and mouse epidermis. A. Immunofluorescence staining for PEDF (green) and DAPI (blue) in the epithelial cells of human expanded skin (n = 9) and control skin (n = 6, ×40 magnification); B. Quantification of PEDF-positive cells per field in human skin (n = 6, mean ± SEM; **P < 0.01); C. Immunofluorescence staining for PEDF (green) and DAPI (blue) in the epithelial cells in mouse expanded skin and control skin (n = 6, ×40 magnification); D. Quantification of PEDF-positive cells per field in mouse skin (n = 6, mean ± SEM; ***P < 0.001). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 3. PEDF accumulated in the subcutaneous exudates of a rat skin expansion model. The quantity of PEDF protein in the subcutaneous exudates obtained at indicated times in expanded and control rats were determined by ELISA assays (n = 6, mean ± SEM; ***P < 0.001).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Control, Enzyme-linked Immunosorbent Assay

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 2. Skin expansion resulted in dermal thinning and PEDF up-regulation in a mouse skin expansion model. A. H&E staining at the 3rd week after expansion, showing thinner dermis in the expanded skin (×40 magnification); B. Quantification of dermal thickness between expanded and control skin (n = 6, mean ± SEM; **P < 0.01); C. The mRNA expression levels of PEDF in skin obtained at the 3rd week compared with the control skin (n = 6, mean

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Staining, Control, Expressing

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 5. In vivo injection of PEDF promoted M1 macrophage polarization. A. A larger number of CD68+/iNOS+ double-positive macrophages were detected in the PEDF injection group compared with PBS treatment. In contrast to PEDF injection, LR antibody injection decreased the number of CD68+/iNOS+ double-positive macrophages; B. Quantification of M1 macrophages in differently treated skin (mean ± SEM; **P < 0.01，*P < 0.05).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: In Vivo, Injection

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 4. Blockage of PEDF receptor rescued dermal thinning in vivo. A. HE staining results showed that recombinant PEDF protein injection decreased the thickness of expanded dermis, while LR blockage rescued dermal thinning of expanded skin; B. Quantification of dermal thickness between differently treated skin (n = 6, mean ± SEM; ***P < 0.001，*P < 0.05).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: In Vivo, Staining, Recombinant, Injection

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 7. Hypoxia induced PEDF expression in HaCaT cells. The mRNA expression levels of PEDF in HaCaT cells under normoxic and hypoxic condi tions were determined by RT-qPCR (mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Quantitative RT-PCR

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 8. PEDF promoted macrophages to polarize towards the M1 subtype under hypoxic conditions. A. FACs analysis of Raw264.7 single-cell sus pensions prepared after treating with PEDF under hypoxic conditions. CD11c+ cells indicate M1 macrophages. B. Quantification of CD11c+ cells under hypoxia (mean ± SEM; **P < 0.01). C. FACS analysis of Raw264.7 single-cell suspensions prepared after treating with PEDF under hypoxic conditions. CD206+ cells indicate M2 macro phages. D. Quantification of CD206+ cells under hypoxic conditions. The mRNA expression of M1 marker genes (E. iNOS, F. TNF-α) and M2 marker genes (G. Arg-1, H. Ym-1) determined by RT-qPCR and normalized to GAPDH mRNA expression. (mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Marker, Quantitative RT-PCR

Journal: Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie

Article Title: Secreted PEDF modulates fibroblast collagen synthesis through M1 macrophage polarization under expanded condition.

doi: 10.1016/j.biopha.2021.111951

Figure Lengend Snippet: Fig. 9. PEDF hindered collagen syn thesis in a macrophage-mediated manner under hypoxic conditions. A, B. Relative mRNA expression of COLI (A) and COLIII (B) in fibroblasts after co-culture with PEDF treated macro phages determined by RT-qPCR (mean

Article Snippet: After deparaffinized and rehydrated in alcohol series, skin sections were kept in a sodium citrate solution and heated in a microwave oven to 95 °C for 20 min before being incubated with a rabbit anti-mouse PEDF primary polyclonal antibody at a 1:30 dilution (Boster Bio-Engineering, Hubei Province, China) at 4 °C overnight.

Techniques: Expressing, Co-Culture Assay, Quantitative RT-PCR

Journal: Molecules

Article Title: Proteome Analysis of Aflibercept Intervention in Experimental Central Retinal Vein Occlusion

doi: 10.3390/molecules27113360

Figure Lengend Snippet: Volcano plot. Log 2 of the ratio aflibercept/NaCl is plotted on the x-axis. On the y-axis, −log p -value refers to the logarithmized p -value from the t -test used to test if a protein was significantly changed. Statistically significantly changed proteins are located above the horizontal line, which denotes a significance level of 0.05. Components of aflibercept are not included in the volcano plot. PEDF: pigment epithelium-derived factor. DNAJ7C: DnaJ homolog subfamily C member 7. AKAP8: A-kinase anchor protein 8. XAB2: Pre-mRNA-splicing factor SYF1. MRPS7: 28S ribosomal protein S7, mitochondrial. RPS18: 40S ribosomal protein S18. RER1: Protein RER1.

Article Snippet: The primary antibodies used were a rat 1 µg/mL monoclonal anti-endoplasmin antibody (MBS439463, MyBioSource, San Diego, CA, USA) and a rabbit polyclonal 2 µg/mL anti-pigment epithelium-derived factor (PEDF) antibody (MBS2027143, MyBioSource, San Diego, CA, USA).

Techniques: Derivative Assay

Journal: The Journal of General Virology

Article Title: Targeting pseudoknots with Cas13b inhibits porcine epidemic diarrhoea virus replication

doi: 10.1099/jgv.0.002071

Figure Lengend Snippet: Sequences of crRNAs designed to target PEDV

Article Snippet: Subsequently, it was incubated with anti-PEDV antibody (MBS560870; MyBiosource) and GAPDH antibody (sc-47724; Santa Cruz Biotechnology) for 2 h at room temperature on a shaker.

Techniques:

Journal: The Journal of General Virology

Article Title: Targeting pseudoknots with Cas13b inhibits porcine epidemic diarrhoea virus replication

doi: 10.1099/jgv.0.002071

Figure Lengend Snippet: Comparison of PEDV variant pseudoknot sequences and crRNA production. ( a ) Alignment of slippery and pseudoknot sequences between SARS-CoV-2 and PEDV. The alignment compares nucleotide sequences, highlighting the slippery site (boxed) and pseudoknot site (grey) between SARS-CoV-2 and PEDV. Identical nucleotides are marked with asterisks (*) and differences with periods (.) below the sequences. Gaps are shown as dashes (-). ( b ) Sequence diversity among PEDV variants, including CV777, OH851 and CKK1-1. The top diagram shows the general genome organization of PEDV. The bottom diagram shows the multiple sequence alignment generated via CLUSTALW that compares the slippery region (boxed) and pseudoknot region (grey) among three PEDV variants. ( c ) CRISPR-Cas13b system targeting the RdRp ) gene of PEDV. The diagram illustrates CRISPR-Cas13b targeting the RdRp gene, indicating the pseudoknot and nonpseudoknot crRNA sites. ( d ) Schematic of the secondary structures of the PEDV slippery and pseudoknot regions with crRNA targeting sites marked. Sequences targeted by crRNAs #1, #2 and #3 are indicated by the red, green and blue bands, respectively.

Article Snippet: Subsequently, it was incubated with anti-PEDV antibody (MBS560870; MyBiosource) and GAPDH antibody (sc-47724; Santa Cruz Biotechnology) for 2 h at room temperature on a shaker.

Techniques: Comparison, Variant Assay, Sequencing, Generated, CRISPR

Journal: The Journal of General Virology

Article Title: Targeting pseudoknots with Cas13b inhibits porcine epidemic diarrhoea virus replication

doi: 10.1099/jgv.0.002071

Figure Lengend Snippet: Potent inhibition of PEDV transmission via pseudoknot-targeting crRNA #3. ( a ) Quantification of PEDV gene expression. Vero cells were transfected with Cas13b mRNA and crRNAs targeting CKK1-1 of the PEDV strain 24 h before infection. After 24 h, RNA was extracted, and PEDV RdRp and membrane gene expression levels were quantified using qRT-PCR. Results were normalized to RNA expression in cells transfected with nontargeting crRNAs. ( b ) Representative images of PEDV protein (green) and DAPI (blue) staining in Vero cells. Viral protein levels from ( a ) were visualized under a fluorescence microscope. Scale bar: 50 µm. ( c ) Quantification of the percentage of PEDV protein-positive cells relative to DAPI-positive cells, tested as in ( b ). Green bars represent PEDV protein-positive cells compared with total cells (DAPI-positive cells). ( d ) Total proteins from the infected cells in ( a ) were analysed using a western blot assay. Intensity ratios of PEDV proteins, normalized to GAPDH, are shown above and below each corresponding protein band. ( e ) Quantification of PEDV titre. Vero cells were infected with the virus from ( a ). Titration was performed using a plaque assay at 4 dpi. The dilution factor multiplied by the plaque count was used to determine the virus titre ( n =3 for each group). Values are means± sem . One-way ANOVA was used for statistical analysis. *** P <0.0001.

Article Snippet: Subsequently, it was incubated with anti-PEDV antibody (MBS560870; MyBiosource) and GAPDH antibody (sc-47724; Santa Cruz Biotechnology) for 2 h at room temperature on a shaker.

Techniques: Inhibition, Transmission Assay, Gene Expression, Transfection, Infection, Membrane, Quantitative RT-PCR, RNA Expression, Staining, Fluorescence, Microscopy, Western Blot, Virus, Titration, Plaque Assay

Journal: The Journal of General Virology

Article Title: Targeting pseudoknots with Cas13b inhibits porcine epidemic diarrhoea virus replication

doi: 10.1099/jgv.0.002071

Figure Lengend Snippet: Optimization of Cas13b and crRNA for enhanced antiviral activity against PEDV in a dose-dependent manner. (a) Quantification of PEDV gene copies. Vero cells were transfected with 0.5 µg of either crRNA #3 or nontarget crRNA and varying amounts of Cas13b mRNA (0.5, 1.0, 1.5 or 2.0 µg). At 24 h post-transfection, cells were infected with PEDV (0.05 m.o.i.). After another 24 h, PEDV RdRp and membrane gene expression levels in total RNA were quantified using qRT-PCR. Results were normalized to RNA expression in cells transfected with nontargeting crRNAs. (b) The same RNA transfection from (a) was followed by a luminescent cell viability assay (ATP assay) 48 h later. Bioluminescence values were normalized to those of untreated cells. (c) Vero cells were transfected with 2 µg of Cas13b mRNA and various amounts of crRNA #3 (0.1, 0.2, 0.5 or 1.0 µg). After 24 h, the cells were infected with PEDV, and 24 h later, PEDV RdRp and membrane gene expression levels were quantified using qRT-PCR. Results were normalized to RNA expression in cells transfected with nontargeting crRNAs. (d) Luminescent cell viability assay (ATP assay) was performed 48 h after transfection, as shown in (c). Bioluminescence values were normalized to those of untreated cells. Values are means± sem . One-way ANOVA was used for statistical analysis. *** P <0.0001.

Article Snippet: Subsequently, it was incubated with anti-PEDV antibody (MBS560870; MyBiosource) and GAPDH antibody (sc-47724; Santa Cruz Biotechnology) for 2 h at room temperature on a shaker.

Techniques: Activity Assay, Transfection, Infection, Membrane, Gene Expression, Quantitative RT-PCR, RNA Expression, Cell Viability Assay, ATP Assay

Journal: The Journal of General Virology

Article Title: Targeting pseudoknots with Cas13b inhibits porcine epidemic diarrhoea virus replication

doi: 10.1099/jgv.0.002071

Figure Lengend Snippet: Inhibition of PEDV transmission via merafloxacin, a pseudoknot formation inhibitor. ( a ) Quantification of PEDV gene copies. Vero cells were infected with PEDV and treated with the indicated merafloxacin concentrations 1 h later. After 24 h, PEDV RdRp gene expression was quantified using qRT-PCR. Results were normalized to RNA expression in cells treated with DMSO. ( b ) Representative immunofluorescence images of PEDV protein (green) and DAPI (blue) staining in Vero cells. Images are from one of three replicates. Scale bars: 50 µm. ( c ) Quantification of the percentage of PEDV protein-positive cells relative to DAPI-positive cells, tested as in ( b ). ( d ) Cytotoxicity quantification following merafloxacin treatment. Vero cells were treated with the indicated concentrations of merafloxacin dissolved in DMSO. After 24 h, a luminescent cell viability assay (ATP assay) was performed. Bioluminescence values were normalized to those of untreated cells. Values are means± sem .

Article Snippet: Subsequently, it was incubated with anti-PEDV antibody (MBS560870; MyBiosource) and GAPDH antibody (sc-47724; Santa Cruz Biotechnology) for 2 h at room temperature on a shaker.

Techniques: Inhibition, Transmission Assay, Infection, Gene Expression, Quantitative RT-PCR, RNA Expression, Immunofluorescence, Staining, Cell Viability Assay, ATP Assay

Journal: Korean Journal of Pathology

Article Title: The Expression of Pigment Epithelium-Derived Factor in Bladder Transitional Cell Carcinoma

doi: 10.4132/KoreanJPathol.2012.46.3.261

Figure Lengend Snippet: Immunohistochemical staining of pigment epithelium-derived factor (PEDF) in normal urothelium (A) and bladder transitional cell carcinoma (TCC) (B-D). The expression of PEDF has a granular pattern in the cytoplasm of normal and neoplastic urothelial cells (B), and some TCCs do not show PEDF expression (C, D).

Article Snippet: This was followed by the incubation of the sections with primary antibody for two hours at room temperature, where the primary antibodies include mouse monoclonal anti-PEDF antibody (1:200, Merck Millipore, Billerica, MA, USA) and anti-CD34 antibody (1:200, Dako, Santa Barbara, CA, USA).

Techniques: Immunohistochemical staining, Staining, Derivative Assay, Expressing

Journal: Korean Journal of Pathology

Article Title: The Expression of Pigment Epithelium-Derived Factor in Bladder Transitional Cell Carcinoma

doi: 10.4132/KoreanJPathol.2012.46.3.261

Figure Lengend Snippet: Immunohistochemical staining of pigment epithelium-derived factor (PEDF) (A, C) and CD34 (B, D) in bladder transitional cell carcinoma (TCC), and the relationship between PEDF expression and microvessel density (MVD) (E). The degree of MVD is significantly higher in TCCs without PEDF expression (A, B) than TCC with PEDF expression (C, D). Bladder TCC shows a significant reciprocal correlation between PEDF expression and MVD (p=0.005) (E).

Article Snippet: This was followed by the incubation of the sections with primary antibody for two hours at room temperature, where the primary antibodies include mouse monoclonal anti-PEDF antibody (1:200, Merck Millipore, Billerica, MA, USA) and anti-CD34 antibody (1:200, Dako, Santa Barbara, CA, USA).

Techniques: Immunohistochemical staining, Staining, Derivative Assay, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Defining a mechanistic link between pigment epithelium–derived factor, docosahexaenoic acid, and corneal nerve regeneration

doi: 10.1074/jbc.M117.801472

Figure Lengend Snippet: Concentration of compounds used in different treatments PEDF was purchased from Bio Products (Middletown, MD). DHA and atglistatin (a PEDF-R inhibitor) were purchased from Cayman Chemical (Ann Arbor, MI). The 44-mer PEDF peptide was prepared by GenWay Biotech, Inc. (San Diego). NPD1 was a kind gift from Dr. Nicos A. Petasis, Loker Hydrocarbon Research Institute, University of Southern California, Los Angeles.

Article Snippet: Eight-μm serial sections were washed with PBS (three times for 5 min), blocked with 10% normal goat serum plus 0.5% Triton X-100 solution in PBS for 1 h, and then incubated at 4 °C overnight with primary rabbit anti-PEDF-R antibody (Cayman, Ann Arbor, MI; 1:250).

Techniques: Concentration Assay, Ex Vivo, Organ Culture