Journal: Molecular and Cellular Biology

Article Title: Deficiency of a Lipid Droplet Protein, Perilipin 5, Suppresses Myocardial Lipid Accumulation, Thereby Preventing Type 1 Diabetes-Induced Heart Malfunction

doi: 10.1128/MCB.00133-14

Figure Lengend Snippet: Levels of Plin and lipase proteins in the heart. (A) Images of immunoblotting of Plin proteins and lipases using total protein extracts from the heart (15 μg of total protein per lane). Coomassie brilliant blue (CBB) staining of the gel was used as a loading control. (B to F) The graphs show the levels of Plin5, Plin2, Plin3, ATGL, and HSL relative to those in saline-treated control WT mice (n = 3 per group). Open bars, WT mice; filled bars, Plin5−/− mice; ND, not detectable. Data are shown as means ± SEM (*, P < 0.05; **, P < 0.01; and ***, P < 0.001).

Article Snippet: Proteins were probed using the following antibodies: Plin5 (developed in-house [ 8 ] and by Progen [GP31]); Plin2 (GP40; Progen); Plin3 (3883; Prosci); ATGL (2138), phospho-p38 (4511), and p38 (9212) (Cell signaling); pan-cadherin (C3678), PKC (P5704), and p47 phox (SAB4502810) (Sigma); glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (sc-25778) and β-tubulin (sc-9104) (Santa Cruz); hormone-sensitive lipase (HSL) (ab45422) and uncoupling protein 3 (UCP3) (ab3477) (Abcam); p67 phox (610912; BD Bioscience); and acyl coenzyme A (CoA) oxidase (Acox), very long-chain acyl-CoA dehydrogenase (VLCAD), and medium-chain acyl-CoA dehydrogenase (MCAD) (gifts from T. Hashimoto and S. Yamaguchi).

Techniques: Western Blot, Staining, Control, Saline

Journal: Nature Communications

Article Title: Chemical reprogramming of fibroblasts into retinal pigment epithelium cells for vision restoration

doi: 10.1038/s41467-025-67104-w

Figure Lengend Snippet: a Representative immunostaining showing that MEF-derived ciRPE cells express ZO-1, Pax6, Rpe65, Mitf, Best1 and Cralbp. Scale bars, 50 μm. b Representative Z-stack confocal micrographs showing ciRPE cells with typical polarized expression of RPE markers. ZO-1 (green) demonstrates apical localization (top), while Best1 (red) shows basolateral localization (bottom). Scale bars, 10 μm. c Representative transmission electron microscopy image of ciRPE cells showing apical microvilli (yellow arrows), melanin granules (red arrows) and tight junctions (black arrows). Scale bars, 1 μm. d Representative confocal micrograph showing phagocytosis of POSs (green) by ciRPEs. The apical sides of ciRPE cells are stained with ZO-1(violet), whereas nuclei are counterstained with DAPI (blue). Scale bars, 50 μm. e Apical and basal secretion of PEDF and VEGF by MEFs, ciRPE, and pRPE cells cultured on Transwells. Each group was compared to the ciRPE group within apical and basal compartments ( n = 6 independent biological samples per group). f Representative morphological images showing dome structures formed by ciRPE cells during in vitro culture. The red arrows indicate the dome morphology observed under different phase-contrast microscopy conditions. Scale bars, 50 μm. g TEER measurements of MEFs, ciRPE, and pRPE cells over time in culture ( n = 6 independent biological samples per group). Data are mean ± SD. One-way ANOVA was used to assess statistical significance. Three independent experiments were performed with similar results and representative results are shown. Source data are provided as a Source Data file.

Article Snippet: Detailed procedures were followed according to the instructions of the PEDF ELISA kit (CUSABIO, CSB-E08820m and CSB-E08818h) and the VEGF ELISA kit (CUSABIO, CSB-E04756m and CSB-E11718h).

Techniques: Immunostaining, Derivative Assay, Expressing, Transmission Assay, Electron Microscopy, Staining, Cell Culture, In Vitro, Microscopy

Journal: Nature Communications

Article Title: Chemical reprogramming of fibroblasts into retinal pigment epithelium cells for vision restoration

doi: 10.1038/s41467-025-67104-w

Figure Lengend Snippet: a Candidate small molecules identified by preliminary screening with scRCF for reprogramming HEFs into OV. b Schematic diagram of the protocol for the reprogramming of HEFs into hciRPE cells, along with representative morphological changes at indicated time points. HM represents the HEF medium, while RM represents the reprogramming medium and DM refers to the differentiation/maturation medium. Scale bar, 300 μm. c FACS purification of reprogrammed BEST1-EGFP + hciRPE cells. d Representative optical microscopy and TEM images of hciRPE cells showing melanin granules (red arrows). Scale bars, 1 μm. e qRT-PCR analysis showing the expression of RPE-associated genes at the indicated time points during reprogramming ( n = 3 independent biological samples per group). f Representative immunostaining analysis showing positive expression of ZO-1, RPE65, MITF and BEST1 in the BEST1-EGFP - HEFs-derived hciRPE cells. Scale bar, 20 μm. g PCA of samples from day 0, day 12, day 24 and day 38 (hciRPE) of celluar reprogramming, and the control primary hRPE cells. h Heatmap showing differentially expressed genes in HEF to hciRPE cell reprogramming samples at indicated time points. The number above heatmap indicates independent biological replicates. Representative genes (left side of the heatmap) and associated GO (right side of the heatmap) for each block are shown. Red and blue indicate upregulated and downregulated genes, respectively. Differential expression was analyzed using the R package limma (v3.58.1) following normalization with edgeR (v4.0.16). Significantly changed genes were defined by |log₂ fold change| > 1.5 and adjusted p < 0.01 (Benjamini-Hochberg correction). i Polarized secretion of VEGF and PEDF from the apical and basal sides of hciRPE cells grown on Transwells ( n = 6 independent biological samples per group). j TEER in hciRPE cells for 30 days. p values indicate comparisons between adjacent time points ( n = 5 independent biological samples per group). Data are mean ± SD. Statistical analyses were performed using one-way ANOVA ( e ) and unpaired, two-tailed Student’s t- test ( i , j ). Three independent experiments were performed with similar results and representative results are shown. Source data are provided as a Source Data file.

Article Snippet: Detailed procedures were followed according to the instructions of the PEDF ELISA kit (CUSABIO, CSB-E08820m and CSB-E08818h) and the VEGF ELISA kit (CUSABIO, CSB-E04756m and CSB-E11718h).

Techniques: Purification, Microscopy, Quantitative RT-PCR, Expressing, Immunostaining, Derivative Assay, Control, Blocking Assay, Quantitative Proteomics, Two Tailed Test

Journal: Stem Cells International

Article Title: Neurotrophic Effect of Adipose Tissue-Derived Stem Cells on Erectile Function Recovery by Pigment Epithelium-Derived Factor Secretion in a Rat Model of Cavernous Nerve Injury

doi: 10.1155/2016/5161248

Figure Lengend Snippet: PEDF in different passages of rat ADSC-conditioned medium. Five samples per passage were analyzed using a rat PEDF ELISA kit. The different passages of the rat ADSCs secreted a detectable level of PEDF in cultured serum-free DMEM/F12 medium. ∗ P < 0.05 compared with the DMEM/F12 medium group only. PEDF = pigment epithelium-derived factor; ADSCs = adipose tissue-derived stem cells; DMEM/F12 = Dulbecco's Modified Eagle's Medium/F12.

Article Snippet: The culture supernatant from the ADSCs was subsequently collected and analyzed using a rat PEDF ELISA kit (CUSABIO, Wuhan, Hubei province, China) according to the manufacturer's instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Derivative Assay, Modification

Journal: Stem Cells International

Article Title: Neurotrophic Effect of Adipose Tissue-Derived Stem Cells on Erectile Function Recovery by Pigment Epithelium-Derived Factor Secretion in a Rat Model of Cavernous Nerve Injury

doi: 10.1155/2016/5161248

Figure Lengend Snippet: Western blotting analyses of PEDF, p-Akt, and p-eNOS protein expression in the rat penile tissues of all groups. (a) Representative Western blot and (b) quantitative analysis identified increased PEDF expression in the bilateral cavernous nerve injury (BCNI) group compared with the sham group at day 14, which recovered to the background level at day 28. ∗ P < 0.05. (c)-(d) Increased PEDF expression in the ADSCs treatment group compared with the other groups at day 28. ∗ P < 0.05. (e)-(f) Increased p-Akt expression in the ADSCs group compared with the other groups at day 28 and increased p-Akt expression in the sham group compared with the bilateral cavernous nerve injury and PBS groups. ∗ P < 0.05 and # P < 0.05. (g-h) Increased p-eNOS expression in the ADSCs treatment group compared with the other groups at day 28. ∗ P < 0.05. The data were normalized to GAPDH protein expression. n = 6 per group. p-Akt = phosphorylated Akt; p-eNOS = phosphorylated endothelial nitric oxide synthase; GAPDH = glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: The culture supernatant from the ADSCs was subsequently collected and analyzed using a rat PEDF ELISA kit (CUSABIO, Wuhan, Hubei province, China) according to the manufacturer's instructions.

Techniques: Western Blot, 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. 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