Journal: Biologics : Targets & Therapy

Article Title: Therapeutic Potential of Compounds with High Affinity to BAG2 in Inhibiting Keloid Disease

doi: 10.2147/BTT.S533286

Figure Lengend Snippet: Inhibition of BAG2 reduced keloid collagen synthesis and deposition. ( A ) The relative gene expression of COL1A1 and TIMP1 in si-BAG2-treated keloid fibroblasts compared to control groups. Gene expression related to collagen synthesis and the inhibition of collagen degradation was significantly reduced following BAG2 inhibition; ( B ) Western blot analysis showing reduced protein expression of BAG2, COL1, COL3, and α-SMA in si-BAG2-treated keloid fibroblasts; ( C ) Masson’s trichrome staining of keloid explants from different groups. Collagen structures in the si-BAG2-treated groups appeared thinner compared to control groups; ( D ) Western blot analysis demonstrates decreasing protein expression of COL1 and COL3 in si-BAG2-treated ex-vivo keloid explants. The reduction in COL1 and COL3 protein expression was statistically significant. Statistical analysis of relative protein expression is provided (n=3). ** P<0.01, **** P<0.0001 .

Article Snippet: KFs were treated with si-BAG2, C16-PAF (Targetmol, # T21547 , 10 nM), or si-BAG2 alone for 24 and 48 hours, along with the si-NC control.

Techniques: Inhibition, Gene Expression, Control, Western Blot, Expressing, Staining, Ex Vivo

Journal: Biologics : Targets & Therapy

Article Title: Therapeutic Potential of Compounds with High Affinity to BAG2 in Inhibiting Keloid Disease

doi: 10.2147/BTT.S533286

Figure Lengend Snippet: Inhibition of BAG2 reduced KF migration and proliferation via the MEK signaling pathway. ( A ) Representative images and bar graph depict the migration of KFs treated with or without si-BAG2 at 0 and 24 hours after scratching (n=3). Scale bar = 500 μm; ( B ) CCK-8 assays were performed on control and si-BAG2-treated KFs, showing a significant reduction in the proliferation of si-BAG2-treated cells (n=3); ( C ) Relative gene expression of TGF-β in si-BAG2-treated and control KFs, with significantly decreased TGF-β expression observed following si-BAG2 treatment (n=3); ( D ) Western blot analysis revealing decreased p-MEK in si-BAG2-treated cells; ( E ) Representative cell cycle profiles of KFs from various groups, assessed by flow cytometry, accompanied by statistical analysis (n=3). *P<0.05, ** P<0.01, ***P<0.001, **** P<0.0001 . KF, keloid fibroblast.

Article Snippet: KFs were treated with si-BAG2, C16-PAF (Targetmol, # T21547 , 10 nM), or si-BAG2 alone for 24 and 48 hours, along with the si-NC control.

Techniques: Inhibition, Migration, CCK-8 Assay, Control, Gene Expression, Expressing, Western Blot, Flow Cytometry

Journal: Cell Death & Disease

Article Title: TNF-alpha-induced microglia activation requires miR-342: impact on NF-kB signaling and neurotoxicity

doi: 10.1038/s41419-020-2626-6

Figure Lengend Snippet: a BAG-1 expression after miR-342 overexpression/inhibition was addressed by western blot. Results were normalized with α-tubulin and compared with the respective controls (mean ± SD, n = 6). To evaluate the involvement of BAG-1 on NF-kB activation, N9 microglia were transfected with a siRNA to silence b or with a plasmid (1ug/mL) to overexpress BAG-1 c . BAG-1 and ph-NF-kB p65 expression levels were evaluated by western blot. Results were normalized with GAPDH and compared with the respective controls (mean ± SD, n = 2–4). Statistical significance: * p < 0.05, Wilcoxon matched-pairs test.

Article Snippet: Overexpression of BAG-1 in N9 microglia was achieved by transiently transfecting the cells with a BAG-1 mammalian expression vector (pCMV6-BAG-1, Origene).

Techniques: Expressing, Over Expression, Inhibition, Western Blot, Activation Assay, Transfection, Plasmid Preparation

Journal: Cell Death & Disease

Article Title: TNF-alpha-induced microglia activation requires miR-342: impact on NF-kB signaling and neurotoxicity

doi: 10.1038/s41419-020-2626-6

Figure Lengend Snippet: We found miR-342 to be upregulated in microglia activated with TNF-α. miR-342 promotes NF-kB activation by inhibiting BAG-1, leading to the overexpression of pro-inflammatory mediators, including TNF-α, in a positive feedback loop, possibly perpetuating microglia activation. Importantly, inhibition of miR-342 attenuated TNF-α-driven microglia activation. Moreover, microglia activation by miR-342 led to increased neurotoxicity with high levels of nitrites being detected in co-cultures supernatants.

Article Snippet: Overexpression of BAG-1 in N9 microglia was achieved by transiently transfecting the cells with a BAG-1 mammalian expression vector (pCMV6-BAG-1, Origene).

Techniques: Activation Assay, Over Expression, Inhibition