Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Hypoxic mesenchymal stem cell-Derived Exosomal circPTP4A2 improves granulosa cell mitochondrial function via YBX1

doi: 10.1007/s00018-025-06067-z

Figure Lengend Snippet: CircPTP4A2 released by H-Exs alleviates mitochondrial damage in GCs. A . QRT-PCR to determine the relative expression levels of circPTP4A in CTX-KGNs co-cultured with N-Exs, H-Exs, and H-Exs-si-circPTP4A2. B-D. Measurement of MDA (B), SOD (C) and GPX (D) activity in cocultured cells. E. Expression level of mtDNA of cocultured cells. F. The ROS level of co-cultured cells was detected by DCFH-DA staining; green for DCFH-DA and blue for nucleus (DAPI) (scale bar = 75 μm). G. SA-β-gal staining showing senescence in co-cultured cells. Senescent cells were stained blue (scale = 200 micrometers) (scale bar = 200 μm). H. The changes of MMP in co-cultured cells were detected by JC-1 staining (scale bar = 75 μm). I. Observation of mitochondrial morphology of co-cultured cells by transmission electron microscopy. (scale bar = 500 nm). J-K. ATP generation capacity (J) and OCR (K) of cocultured cells. L. The binding mode of the complex YBX1 protein with circPTP4A2 after 100ns MD simulation. M. The RMSD of circPTP4A2 with YBX1 protein. N. The RMSF of circPTP4A2 with YBX1 protein. O. The Rg of circPTP4A2 with YBX1 protein. P. The SASA of circPTP4A2 with YBX1 protein. Q. The hydrogen bond number of circPTP4A2 with YBX1 protein. R. RIP assay to measure circPTP4A2 expression in YBX1-overexpressing HEK 293 T cells. S-T . QRT-PCR analysis of circPTP4A2 expression after YBX1 knockdown or overexpression. U. Relative YBX1 mRNA and protein levels after circPTP4A2 knockdown. V-W. After CHX reagent treatment, WB assay was performed to detect YBX1 protein level after circPTP4A2 knockdown.Protein quantification of YBX1 (W). X. After MG132 reagent treatment, WB assay was performed to detect YBX1 protein level after circPTP4A2 knockdown

Article Snippet: Molecular dynamics simulations revealed that circPTP4A2 (U-331, A-326, A-146, G-144, G-145, G-333, C-147, U-327, and U-143) can interact with YBX1 (ARG-69, LYS-137, ASP-83, ALA-120, LYS-118, ASN-70, TRP-65, TYR-138, and LYS-52).The results of this investigation elucidated the interaction between circPTP4A2 and YBX1, revealing that circPTP4A2 not only binds to YBX1 but also increases its stability by protecting it from proteasomal degradation.

Techniques: Quantitative RT-PCR, Expressing, Cell Culture, Activity Assay, Staining, Transmission Assay, Electron Microscopy, Binding Assay, Knockdown, Over Expression

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Hypoxic mesenchymal stem cell-Derived Exosomal circPTP4A2 improves granulosa cell mitochondrial function via YBX1

doi: 10.1007/s00018-025-06067-z

Figure Lengend Snippet: Overexpression of YBX1 improved the oxidative damage induced by circPTP4A2 Silencing. A-C. Measurement of MDA (A), SOD (B) and GPX (C) activity in co-cultured cells. D. The ROS level of co-cultured cells was detected by DCFH-DA staining; green for DCFH-DA and blue for nucleus (DAPI) (scale bar = 75 μm). E. SA-β-gal staining showing senescence in co-cultured cells. Senescent cells were stained blue (scale = 200 micrometers) (scale bar = 200 μm). F. The changes of MMP in co-cultured cells were detected by JC-1 staining (scale bar = 75 μm). G. Observation of mitochondrial morphology of co-cultured cells by transmission electron microscopy. (scale bar = 500 nm). H-J. Relative level of mtDNA (H), ATP generation capacity (I) and OCR (J) of cocultured cells (&: si-NC vs. si-circPTP4A2; #: si-circPTP4A2 vs. si-circPTP4A2 + YBX1)

Article Snippet: Molecular dynamics simulations revealed that circPTP4A2 (U-331, A-326, A-146, G-144, G-145, G-333, C-147, U-327, and U-143) can interact with YBX1 (ARG-69, LYS-137, ASP-83, ALA-120, LYS-118, ASN-70, TRP-65, TYR-138, and LYS-52).The results of this investigation elucidated the interaction between circPTP4A2 and YBX1, revealing that circPTP4A2 not only binds to YBX1 but also increases its stability by protecting it from proteasomal degradation.

Techniques: Over Expression, Activity Assay, Cell Culture, Staining, Transmission Assay, Electron Microscopy

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Hypoxic mesenchymal stem cell-Derived Exosomal circPTP4A2 improves granulosa cell mitochondrial function via YBX1

doi: 10.1007/s00018-025-06067-z

Figure Lengend Snippet: CircPTP4A2 Mitigates Ovarian Dysfunction in POI Rats via Interaction with YBX1. A . Immunofluorescent images stained for FSHR, YBX1 of ovaries in each group. Scale bar, 20 μm. Green: FSHR stained GCs. magenta: YBX1 staining. Blue: DAPI-labeled nucleus. B. Schematic diagram of the research mechanism

Article Snippet: Molecular dynamics simulations revealed that circPTP4A2 (U-331, A-326, A-146, G-144, G-145, G-333, C-147, U-327, and U-143) can interact with YBX1 (ARG-69, LYS-137, ASP-83, ALA-120, LYS-118, ASN-70, TRP-65, TYR-138, and LYS-52).The results of this investigation elucidated the interaction between circPTP4A2 and YBX1, revealing that circPTP4A2 not only binds to YBX1 but also increases its stability by protecting it from proteasomal degradation.

Techniques: Staining, Labeling