Journal: Journal of Ovarian Research

Article Title: Trophoblast cell surface antigen 2 (TROP2) from ascitic extracellular vesicles drives peritoneal metastasis of ovarian cancer by mesothelial-to-mesenchymal transition

doi: 10.1186/s13048-025-01845-6

Figure Lengend Snippet: Mesothelial-mesenchymal transition of peritoneal mesothelial cells induced by ascites-derived EVs. A TEM analysis of EVs. B NTA analysis of EVs. C Western blot analysis of EVs markers. D Dil-labeled EVs were incubated with HMrSV5 cells for 24 h and then observed under a fluorescence microscope (Dil in red and DAPI in blue). HMrSV5 cells were incubated with EVs derived from Cancer EVs and Normal EVs, respectively. The PBS group and the TGF-β1 (0.5 ng/mL) + IL-1β (2.5 ng/mL) group served as negative and positive controls, respectively. E - H . The protein levels of E-Cadherin, N-Cadherin and Vimentin in HMrSV5 cells incubated with Cancer EVs and Normal EVs. I - J . The effect of HMrSV5 cells incubated with Cancer EVs and Normal EVs migration analyzed by Transwell assay. K - M . Representative images depicting the adhesion of CMTPX-labelled SKOV3 (red) and OVCAR3 (red) cells to HMrSV5 cells (blue) are shown and adhered cells were quantified. Data are representative of at least three independent experiments and are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: D-Luciferin potassium salt was obtained from Yeasen (China), recombinant human TGF-β1 and human IL-1β were from R&D Systems (USA), and quinazoline (QNZ) was purchased from MCE (USA).

Techniques: Derivative Assay, Western Blot, Labeling, Incubation, Fluorescence, Microscopy, Migration, Transwell Assay

Journal: bioRxiv

Article Title: Macrophage Migration Inhibitory Factor (MIF)-CD74 Signaling Pathway Mediates Trabecular Meshwork Dysfunction in Glaucoma

doi: 10.64898/2026.03.18.712673

Figure Lengend Snippet: Wild-type C57BL/6 mice (n=12/group) were injected intravitreally with either LV-TGF-β2 or LV-null vectors. ( A ) Weekly IOP measurements demonstrate significant and sustained IOP elevation in LV-TGF-β2-injected mice compared with LV-null controls. ( B ) Immunofluorescence staining of anterior segment cross-sections for α-SMA and MIF shows co-localization of MIF with the TM marker α-SMA, indicating increased MIF expression in the TM of LV-TGF-β2 injected mice. ( C-E ) qPCR ( C ) and western blot ( D, E ) analyses of anterior segment tissues show upregulation of MIF and CD74 accompanied by suppression of Blimp-1 in LV-TGF-β2-injected mice. Increased expression of profibrotic markers, including α-SMA, Fn-1, and MYOC, was also evident in LV-TGF-β2-injected mice. Western blot bands were quantified by densitometric analysis using ImageJ and normalized to the housekeeping control β-actin, and protein levels are presented as relative expression (AU) ( E ). Data represent mean ± SD. **p<0.005, ***p<0.0005, and ****p<0.0001. Two-way ANOVA with Tukey’s multiple-comparison test (A), unpaired t-test (C, E).

Article Snippet: Recombinant human TGF-β2 (302-B2-010/CF) protein was purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection, Immunofluorescence, Staining, Marker, Expressing, Western Blot, Control, Comparison

Journal: bioRxiv

Article Title: Macrophage Migration Inhibitory Factor (MIF)-CD74 Signaling Pathway Mediates Trabecular Meshwork Dysfunction in Glaucoma

doi: 10.64898/2026.03.18.712673

Figure Lengend Snippet: Primary HTMCs (n=3) were challenged with TGF-β2 (10ng/mL), rMIF (100ng/mL), or pro-inflammatory cytokine pool (CP; TNF-α, IL-1β, IL-6, and IL-17 100ng/mL) for 24h. (A) qPCR analysis shows significant upregulation of MIF and CD74 transcripts accompanied by reduced Blimp-1 expression in response to glaucomatous stressors. ( B ) Immunofluorescence staining demonstrates increased MIF expression following TGF-β2 or CP treatment. (C) Western blot analysis confirms induction of MIF and CD74 with concurrent suppression of Blimp-1 at the protein level. Data represent mean ± SD. **p<0.005, ***p<0.0005, and ***p<0.0001, One-way ANOVA with Dunnett’s multiple-comparison test.

Article Snippet: Recombinant human TGF-β2 (302-B2-010/CF) protein was purchased from R&D Systems (Minneapolis, MN).

Techniques: Expressing, Immunofluorescence, Staining, Western Blot, Comparison

Journal: bioRxiv

Article Title: Macrophage Migration Inhibitory Factor (MIF)-CD74 Signaling Pathway Mediates Trabecular Meshwork Dysfunction in Glaucoma

doi: 10.64898/2026.03.18.712673

Figure Lengend Snippet: Primary HTMCs (n=3) were challenged with rMIF (100 ng/mL) in the presence or absence of MIF inhibitor 4-IPP (100µM) or the metabolites Agm or Thia (100 ng/mL) for 24h. Cells treated with TGF-β2 (10ng/mL) served as a positive control. (A-B) Western blot analysis confirms activation of the RhoA/ROCK/pMLC signaling pathway in response to MIF treatment, which is significantly attenuated by MIF inhibition with 4-IPP, Agm, or Thia ( A ). Western blot bands were quantified by densitometric analysis using ImageJ and normalized to the housekeeping control β-actin, and protein levels are presented as relative expression (AU) ( B ). (C) Immunofluorescence analysis shows that MIF induces pMLC staining, comparable to that observed with TGF-β2 stimulation, which is significantly attenuated by MIF inhibition with 4-IPP, Agm, or Thia. Data represent mean ± SD. *p<0.05, **p<0.005, ***p<0.0005, and ****p<0.0001, One-way ANOVA with Dunnett’s multiple-comparison test.

Article Snippet: Recombinant human TGF-β2 (302-B2-010/CF) protein was purchased from R&D Systems (Minneapolis, MN).

Techniques: Positive Control, Western Blot, Activation Assay, Inhibition, Control, Expressing, Immunofluorescence, Staining, Comparison

Journal: bioRxiv

Article Title: Macrophage Migration Inhibitory Factor (MIF)-CD74 Signaling Pathway Mediates Trabecular Meshwork Dysfunction in Glaucoma

doi: 10.64898/2026.03.18.712673

Figure Lengend Snippet: OHT was induced in WT C57BL/6 mice (n=12) by intravitreal injection of LV-TGF-β2. LV-null-injected mcie served as controls. Following OHT induction, extracellular vesicle-loaded agmatine (EV-Agm; 0.1 µg/eye) was applied topically once daily. (A) Weekly IOP measurements over 9 weeks show that EV-Agm treatment significantly reduces IOP compared with untreated LV-TGF-β2-injected mice. (B) qPCR analysis of anterior segment tissue demonstrates that EV-Agm significantly reduces expression of Mif, Cd74, pro-inflammatory cytokines ( Il-1 β , Il-6 ), and cytoskeletal/ECM markers (α -Sma, Myoc, Fn-1 ), while restoring Blimp-1 expression compared with untreated LV-TGF-β2-injected mice. *p<0.05, **p<0.005, ###,***p<0.0005, and ####,****p<0.0001. Two-way ANOVA with Tukey’s multiple comparison (A), one-way ANOVA with Tukey’s multiple comparison (B). In panel A, * denotes LV-null vs. LV-TGF-β2, and # denotes LV-TGF-β2 vs. LV-TGF-β2+EV-Agm comparisons.

Article Snippet: Recombinant human TGF-β2 (302-B2-010/CF) protein was purchased from R&D Systems (Minneapolis, MN).

Techniques: Injection, Expressing, Comparison