Journal: Science Advances

Article Title: Intracellular enzymatic reducing systems control receptor tyrosine kinase signaling via PTP1B

doi: 10.1126/sciadv.adv5362

Figure Lengend Snippet: ( A ) MEF cells reconstituted with hPTP1B were grown on coverslips, serum starved, and stimulated with PDGF (50 ng/ml) for 4 min, and subsequently treated with 10 μM PDGFR inhibitor (PDGFRi) for 5, 10, and 20 min. Cells were fixed, permeabilized at the indicated times, and subsequently evaluated with primary antibodies from different species for hPTP1B (mouse) and GLRX1 (goat) to reveal protein-protein interactions in situ using PLA (red spots). Nuclei were labelled with 4′,6-diamidino-2-phenylindole (DAPI; blue). ( B ) Visualization and quantification were performed using fluorescent microscopy and ImageJ. Statistical differences in signal quantification are indicated in the bar graph (* P < 0.05; ** P < 0.01); n ≥ 3. Error bars denote SEM. ( C ) MEF cells reconstituted with hPTP1B were grown on coverslips, serum starved, and stimulated with PDGF (50 ng/ml) for 4 min or with 100 μM H 2 O 2 for 10 min, with and without catalase pretreatment (2 hours; 50 U/ml) as indicated. The images display results from one representative experiment, out of three independent experiments. Scale bars, 30 μm. Data were analyzed using one-way analysis of variance followed by Bonferroni post hoc tests for multiple comparisons with GraphPad Prism.

Article Snippet: After overnight starvation, the cells were stimulated with either EGF or PDGF (R&D Systems, catalog no. 220-BB) ligand for 4 min.

Techniques: Protein-Protein interactions, In Situ, Microscopy

Journal: Advanced Science

Article Title: Endothelial PDGF Signaling Dysregulation Impairs Testicular Interstitial Homeostasis in Diabetes

doi: 10.1002/advs.202520114

Figure Lengend Snippet: PDGF‐BB supplementation ameliorates interstitial cell dysfunction. (A) Schematic diagram of ex vivo culture of human testicular tissue derived from diabetic patients (n = 6). Tissue fragments were cultured with or without PDGF‐BB supplementation (100 ng/mL) for 3 days, followed by ELISA, RT‐PCR, and immunofluorescence validation. This plot was created with BioRender.com. (B) Representative immunofluorescence staining of JUND (magenta) and CYP17A1 (green) in ex vivo cultured diabetic human testicular tissue with or without PDGF‐BB treatment (100 ng/mL). Nuclei are counterstained with DAPI (blue). Insets show magnified views. Scale bars, 50 µm; 10 µm (insets). (C) Bar graph quantifying the proportion of JUND + CYP17A1 + LCs in ex vivo cultured diabetic human testicular tissue with or without PDGF‐BB supplementation (100 ng/mL). Bars represent the mean ± SD. n = 3 independent biological replicates per group. **** p < 0.0001. (D) Representative immunofluorescence staining of MCL1 (magenta) and CYP11A1 (green) in ex vivo cultured diabetic human testicular tissue with or without PDGF‐BB treatment (100 ng/mL). Nuclei are counterstained with DAPI (blue). Insets show magnified views. Scale bars, 50 µm; 10 µm (insets). (E) Bar graph quantifying the proportion of MCL1 + CYP17A1 + LCs in ex vivo cultured diabetic human testicular tissue with or without PDGF‐BB supplementation (100 ng/mL). Bars represent the mean ± SD. n = 3 independent biological replicates per group. **** p < 0.0001. (F) Representative immunofluorescence staining of INSL3 (magenta), Collagen III (green), and TUNEL (yellow) in ex vivo cultured diabetic human testicular tissue with or without PDGF‐BB (100 ng/mL). Nuclei are counterstained with DAPI (blue). Insets show magnified views. Scale bars, 50 µm; 10 µm (insets). (G) Bar graph showing testosterone concentrations measured by ELISA in supernatants from ex vivo cultured diabetic human testicular tissue with or without PDGF‐BB supplementation (100 ng/mL). Bars represent mean ± SD. * p < 0.05. (H‐J) RT‐PCR analysis of MYH11, MYL9 , and MYLK expression in ex vivo cultured diabetic human testicular tissue with or without PDGF‐BB supplementation (100 ng/mL). Bars represent mean ± SD. n = 3 independent biological replicates per group. **** p < 0.0001, * p < 0.05. Statistical significance was determined using the two‐tailed Student's t‐test (C, E, G, H, I, J).

Article Snippet: Where indicated, recombinant human PDGF‐BB (R&D Systems, 220‐BB‐010) was added at a final concentration of 100 ng/mL.

Techniques: Ex Vivo, Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Biomarker Discovery, Staining, TUNEL Assay, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: Endothelial PDGF Signaling Dysregulation Impairs Testicular Interstitial Homeostasis in Diabetes

doi: 10.1002/advs.202520114

Figure Lengend Snippet: TEC‐derived PDGF signaling disruption impairs interstitial homeostasis and is partially restored by PDGF‐BB supplementation. In healthy testes, TECs secrete PDGFB, which engages PDGFRB on LCs and TPCs to maintain interstitial homeostasis. Under diabetic conditions, TECs undergo EndMT and show a marked reduction in PDGFB expression. Together with decreased PDGFRB levels in LCs and TPCs, this collapse of the ligand‐receptor axis disrupts intercellular communication and contributes to interstitial dysfunction. Specifically, LCs exhibit profound structural and functional alterations, including excessive ECM deposition, increased apoptosis resulting from inactivation of the JUND‐MCL1 survival pathway, and diminished testosterone production. Meanwhile, TPCs display impaired contractility and thinning of the basement membrane, further destabilizing the interstitial niche. Importantly, supplementation with exogenous PDGF‐BB partially restores this paracrine support, enhancing LC survival and steroidogenic activity and improving TPC contractility, thereby alleviating diabetes‐induced testicular dysfunction. This plot was created with BioRender.com.

Article Snippet: Where indicated, recombinant human PDGF‐BB (R&D Systems, 220‐BB‐010) was added at a final concentration of 100 ng/mL.

Techniques: Derivative Assay, Disruption, Expressing, Functional Assay, Membrane, Activity Assay