Journal: Oncogene

Article Title: Disruption of androgen receptor-cofactor interactions by the RNA-binding protein FUS/TLS alters androgen signalling in prostate cancer

doi: 10.1038/s41388-026-03682-3

Figure Lengend Snippet: a COS-1 were transfected with a Gal4 luciferase reporter and β-galactosidase expression plasmid, expression plasmids for Gal4 fused full-length of truncations of FUS and p300, SRC1 or ARA70. b COS-1 were transfected with plasmids encoding the AR, SRC-1, FUS and β-galactosidase, and an AR-responsive luciferase reporter. Luciferase activity was expressed as a % of AR activity in the presence of mibolerone (MIB) and absence of SRC1 and FUS. Mean ± 1SE of 3 independent repeats in duplicate. c Immunoblotting was performed to confirm successful expression of the AR, SRC1 and FUS. d LNCaP FUS were treated ± MIB ± DOX and chromatin immunoprecipitation performed using antibodies specific to the AR, SRC1, RNA Pol II, FUS, or IgG control. Enrichment of the KLK3 enhancer was analysed using qPCR. Mean ± 1SE of 3 independent repeats. ANOVA * p < 0.05, ** p < 0.005, *** p < 0.0005.

Article Snippet: Immunoblotting was performed as previously described [ ] using the following primary antibodies: AR (N-20), FUS (4H11), EWS (G-5) antibodies were from Santa Cruz (CA, USA), SRC-1 antibody (21952-1-AP) was from Proteintech (IL, USA).

Techniques: Transfection, Luciferase, Expressing, Plasmid Preparation, Activity Assay, Western Blot, Chromatin Immunoprecipitation, Control

Journal: Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas

Article Title: Periodic mechanical stress activates MEK1/2-ERK1/2 mitogenic signals in rat chondrocytes through Src and PLCγ1.

doi: 10.1590/s0100-879x2011007500150

Figure Lengend Snippet: Figure 4. Src shRNA and ERK1/2 shRNA inhibit periodic mechanical stress-induced chondrocyte proliferation and matrix synthesis. A, Chondrocytes were transfected with shRNA targeted to Src or ERK1/2 or with non-targeting (NT) sequences prior to lysis and Western blot for Src and ERK1/2 protein. Either the Src or ERK1/2 shRNA sequence achieved about a 50% reduction in Src or ERK1/2 protein levels. After pretreatment with control shRNA or Src shRNA and ERK1/2 shRNA, rat chondrocytes were cultured for 3 days under static conditions or conditions of periodic mechanical stress 8 h per day prior to proliferation studies. Cell proliferation was analyzed by direct cell counting (B) and CCK-8 assay (C). Rat chondrocytes were cultured in vitro for 8 h with static conditions or periodic mechanical stress. Aggrecan (D) and type II collagen (E) gene expression were measured by quantitative real-time PCR. Cell proliferation and matrix synthesis in the inhibitor pretreatment groups was significantly decreased relative to control in chondrocytes in response to periodic mechanical stress (N = 5, *P < 0.05, Student unpaired t-test).

Article Snippet: Src shRNA (r) Lentiviral Particles, ERK1 shRNA (r) Lentiviral Particles, ERK2 shRNA (r) Lentiviral Particles, Control shRNA Lentiviral Particles, and Polybrene were supplied by Santa Cruz.

Techniques: shRNA, Transfection, Lysis, Western Blot, Sequencing, Control, Cell Culture, Cell Counting, CCK-8 Assay, In Vitro, Gene Expression, Real-time Polymerase Chain Reaction

Journal: Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas

Article Title: Periodic mechanical stress activates MEK1/2-ERK1/2 mitogenic signals in rat chondrocytes through Src and PLCγ1.

doi: 10.1590/s0100-879x2011007500150

Figure Lengend Snippet: Figure 6. Effect of PP2 and Src shRNA on the expression and phosphorylation of Src, PLCγ1, MEK1/2, and ERK1/2 under con ditions of periodic mechanical stress. After pretreatment with DMSO or Src inhibitor (PP2), or Src shRNA or non-targeting (NT) sequence, rat chondrocytes were cultured in vitro for 1 h under static conditions or conditions of period ic mechanical stress. The expres sion and phosphorylation levels of Src (A), PLCγ1 (B), MEK1/2 (C), and ERK1/2 (D) were detected by Western blotting. For each West ern blot, the corresponding total amount of each signaling protein served as a control. The images above the histogram are repre sentative results of several inde pendent Western blots. The phos phorylation levels of Src-Tyr418 in the PP2 groups were significantly reduced compared to those in the control groups in chondrocytes in response to periodic mechanical stress (N = 5, *P < 0.05, Student unpaired t-test). The phospho rylation levels of PLCγ1-Tyr783, MEK-Ser217/221 and ERK1/2- Thr202/Tyr204 in the PP2 and Src shRNA groups were significantly reduced compared to those in the control groups in chondrocytes in response to periodic mechanical stress (N = 5, *P < 0.05, Student unpaired t-test).

Article Snippet: Src shRNA (r) Lentiviral Particles, ERK1 shRNA (r) Lentiviral Particles, ERK2 shRNA (r) Lentiviral Particles, Control shRNA Lentiviral Particles, and Polybrene were supplied by Santa Cruz.

Techniques: shRNA, Expressing, Phospho-proteomics, Sequencing, Cell Culture, In Vitro, Western Blot, Control

Journal: BMC Pharmacology & Toxicology

Article Title: Investigating the mechanisms by which bisphenol A affects osteoarthritis through a novel network toxicology framework and experimental validation

doi: 10.1186/s40360-026-01108-0

Figure Lengend Snippet: Experimental validation. ( A , C , E , G , I and K ) Relative expression of SRC, ESR1, EGFR, PTGS2, PPARG and HSP90AA1 via RT-qPCR. ( B , D , F , H , J and L ) Typical pictures and scores of the relative expression of SRC, ESR1, EGFR, PTGS2, PPARG and HSP90AA1 via IHC; 20×, Scale bar: 100 μm; 40×, Scale bar: 50 μm. ** P < 0.01, and *** P < 0.001

Article Snippet: The sections were incubated overnight at 4 °C with the following primary antibodies (all from Proteintech, diluted in PBS): SRC (1:200, 60315-1-Ig), ESR1 (1:200, 21244-1-AP), EGFR (1:1000, 18986-1-AP), PTGS2 (1:1000, 66351-1-Ig), PPARG (1:400, 16643-1-AP), and HSP90AA1 (1:400000, 13171-1-AP).

Techniques: Biomarker Discovery, Expressing, Quantitative RT-PCR