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