Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: DJ-1 promotes necroptosis of chondrocytes by the PTEN/PI3K-AKT signaling pathway

doi: 10.1007/s00018-026-06122-3

Figure Lengend Snippet: DJ-1 Promotes TSZ + LPS-Induced Dysregulation of Chondrocyte Matrix Metabolism. ( A , B ) The protein levels of PTEN, DJ-1, Aggrecan, Collagen II, and MMP3, MMP13 were detected by western blot in human primary chondrocytes overexpressing HA-DJ-1. Corresponding total protein and β-actin were used as loading controls. ( C ) The mRNA levels of Aggrecan, Collagen II, and MMP3, MMP13 were detected by qRT-PCR in human primary chondrocytes overexpressing HA-DJ-1. ( D , E ) Immunofluorescence staining was performed to analyze Aggrecan and MMP13 expression in human primary chondrocytes overexpressing HA-DJ-1. Aggrecan (green), MMP13(red), DAPI (blue), (Scale bar: 20 μm). ( F , G ) Western blot analysis of PTEN, DJ-1, Aggrecan, Collagen II, MMP3, and MMP13 protein in different groups with pretreatment of PTEN activator Oroxin B (0.5µM, MCE). Corresponding total protein and β-actin were used as loading controls. The n values are all biological replicates. Data are presented as mean ± SD of triplicates. Statistical significance is indicated by **** P < 0.0001, *** P < 0.001, ** P < 0.01, * P < 0.05, and ns : not significant ( n = 3 each group). Data were compared using one-way analysis of variance

Article Snippet: Aggrecan (green), MMP13(red), DAPI (blue), (Scale bar: 20 μm). ( F , G ) Western blot analysis of PTEN, DJ-1, Aggrecan, Collagen II, MMP3, and MMP13 protein in different groups with pretreatment of PTEN activator Oroxin B (0.5μM, MCE).

Techniques: Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining, Expressing

Journal: bioRxiv

Article Title: Matrix metalloproteinases proteolyze RAB proteins and contribute to cisplatin-induced ototoxicity

doi: 10.64898/2026.02.28.708770

Figure Lengend Snippet: Gelatin zymography (A, C, E) and corresponding quantitative analysis (B, D, F, G) of MMP-2 and MMP-9 activities in the conditioned media of HEI-OC1 cells following 6 h (A, B) and 48 h (C, D) of exposure to 20 μM cisplatin. Cytosolic MMP-2 and MMP-9 activities were measured following 4 h and 6 h of treatment with 20 μM cisplatin (E-G). Standard (Std) derives from conditioned medium from HT-1080 cells. H) mRNA expression of Ifit-1 was measured using RT-qPCR following a 24h exposure to 20 μM cisplatin in the presence or absence of ARP-100 (5 μM), compared to the untreated control (nil), and normalized to B2M expression. The data are represented as the mean ± SD of three independent experiments. Statistical significance was determined using one-way ANOVA followed by Dunnett’s post-hoc test (* p < 0.05 and ** p <0.01).

Article Snippet: For overexpression studies, the pCMV3-C-FLAG vector encoding human MMP-2 ORF (HG10082-CF, Sino Biological) was transfected into HEI-OC1 using jetPRIME Reagent (Polyplus) according to the manufacturer’s protocol.

Techniques: Zymography, Expressing, Quantitative RT-PCR, Control

Journal: bioRxiv

Article Title: Matrix metalloproteinases proteolyze RAB proteins and contribute to cisplatin-induced ototoxicity

doi: 10.64898/2026.02.28.708770

Figure Lengend Snippet: mRNA expression of N-terminal truncated (NTT)- Mmp-2 (A), full-length (FL)- Mmp-2 (B), and Mmp-9 (C) was measured using RT-qPCR following exposure to 20 μM cisplatin for various timepoints, compared to the untreated control (0), and normalized to B2M expression. D) Immunofluorescence staining of HEI-OC1 cells treated with 20 μM cisplatin for 6 h or untreated cells. Staining was performed using an antibody specific to MMP-2, followed by detection with a secondary antibody conjugated to Alexa Fluor 647 (red). The zoomed-in boxes indicate nuclei with an increased red signal (magenta). The data are represented as the mean ± SD of three independent experiments. Statistical significance was determined using one-way ANOVA followed by Dunnett’s post-hoc test (* p < 0.05).

Article Snippet: For overexpression studies, the pCMV3-C-FLAG vector encoding human MMP-2 ORF (HG10082-CF, Sino Biological) was transfected into HEI-OC1 using jetPRIME Reagent (Polyplus) according to the manufacturer’s protocol.

Techniques: Expressing, Quantitative RT-PCR, Control, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Matrix metalloproteinases proteolyze RAB proteins and contribute to cisplatin-induced ototoxicity

doi: 10.64898/2026.02.28.708770

Figure Lengend Snippet: (A) IL-6 secretion in cells transfected with either an empty vector (EV) or an Mmp-2 -expressing vector and treated with 100 μM cisplatin or left untreated (nil) for 24 h. (B, C) IL-6 secretion in cells pre-treated for 2 h with the vehicle or 5 μM ARP-100 (B) or ONO-4817 (C), followed by treatment with 20 μM cisplatin for 24 h. (D) IL-6 secretion in cells transfected with non-targeting siRNA ( siNT ), Mmp-9 -targeting siRNA ( siMmp-9 ), and exposed to 20 μM cisplatin for 24 h. The data are represented as the mean ± SD of three independent experiments. Statistical significance was determined using one-way ANOVA followed by Dunnett’s post-hoc test (* p < 0.05).

Article Snippet: For overexpression studies, the pCMV3-C-FLAG vector encoding human MMP-2 ORF (HG10082-CF, Sino Biological) was transfected into HEI-OC1 using jetPRIME Reagent (Polyplus) according to the manufacturer’s protocol.

Techniques: Transfection, Plasmid Preparation, Expressing

Journal: bioRxiv

Article Title: Matrix metalloproteinases proteolyze RAB proteins and contribute to cisplatin-induced ototoxicity

doi: 10.64898/2026.02.28.708770

Figure Lengend Snippet: A, B) Immunoblot of in vitro time course proteolysis of recombinant RAB9A at 37 °C with 100 ng MMP-2 (A) or for 30 minutes in the presence or absence of MMP-2-preferring inhibitors (B). C) In silico analysis of the top 10 MMP-2 cleavage sites in the structure of mouse RAB9A according to ProsperousPlus. D) Crystal structure of mouse RAB9A showing the predicted cleavage sites (green balls).

Article Snippet: For overexpression studies, the pCMV3-C-FLAG vector encoding human MMP-2 ORF (HG10082-CF, Sino Biological) was transfected into HEI-OC1 using jetPRIME Reagent (Polyplus) according to the manufacturer’s protocol.

Techniques: Western Blot, In Vitro, Recombinant, In Silico