protein extraction kit  (Keygen Biotech)

Journal: Journal of Advanced Research

Article Title: Spike RBD drives sustained Parkinson’s disease progression via microglia-neuron crosstalk-mediated RTP801 upregulation

doi: 10.1016/j.jare.2025.07.060

Figure Lengend Snippet: RBD exacerbates degeneration of dopaminergic neurons and αSyn aggregation. (A) Representative images of TH immunostaining in the SNc and striatum and quantitative assessment (all values were normalized relative to CSGS 1w + Vehicle group), scale bar = 500 μm, n = 4. (B) Western blot analysis of TH expression in SNc and striatum and quantitative analysis (n = 4). (C) Representative image of Lewy bodies of SNc and quantitative analysis of LB signal intensity (n = 4), scale bar = 50 μm. (D-E) Colocalization of p-αSyn with αSyn in the SNc, quantitative analysis of p-αSyn signal intensity (n = 4), scale bar = 5 μm. (F-G) Colocalization of n-αSyn with αSyn in the SNc, quantitative analysis of n-αSyn signal intensity (n = 4), scale bar = 5 μm. (H) ThS staining quantifies aggregates in the SNc, including ThS signal density and the proportion of cells inclusive ThS + puncta (n = 4), scale bar = 5 μm. (I) Western blot detection of αSyn aggregation using the 5G4 antibody, with quantitative analysis confirming significant changes (n = 4). ( J ) Analysis of αSyn partitioning in soluble and insoluble brain fractions by western blot, with quantitative data indicating significant alterations (n = 4). Data are expressed as means ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant.

Article Snippet: Soluble and insoluble protein fractions were isolated using a commercially accessible Soluble & Insoluble Protein Extraction Kit (Sangon Biotech, Shanghai, China).

Techniques: Immunostaining, Western Blot, Expressing, Staining

Journal: Journal of Advanced Research

Article Title: Spike RBD drives sustained Parkinson’s disease progression via microglia-neuron crosstalk-mediated RTP801 upregulation

doi: 10.1016/j.jare.2025.07.060

Figure Lengend Snippet: RTP801 is critical for PD deterioration induced by RBD. (A) Schematic of the αSyn A53T + ; RTP801 −/− mice experiment. (B) Assessment of olfactory dysfunction was conducted using the BFPT and the visible BFPT. (C) A modified open field test with nuts or paprika quantified olfactory acuity. (D-F) Motor coordination and balance were evaluated using the rotarod (D) , pole (E) , and beam walking (F) tests. ( G ) Forelimb grip strength was gauged through the grip strength test. ( H ) The open field test appraised anxiety-related behaviors, tracking total distance traveled, center zone activity. (I-J) Recognition memory was assessed using Y maze test and NORT. Alternation index (% of total triplet arm entries) was quantified in (I) . New object index was quantified in ( J ) (n = 6). (K) Representative images of TH immunostaining in the SNc and striatum and quantitative assessment (all values were normalized relative to αSyn A53T group), scale bar = 500 μm, n = 4. (L) Western blot analysis of TH expression in SNc and striatum and quantitative analysis (n = 4). (M) Colocalization of RTP801 with TH, scale bar = 200 μm. (N) ThS staining quantifies aggregates in the SNc, including ThS signal density, and the proportion of cells inclusive ThS + puncta (n = 4), scale bar = 5 μm. (O) Western blot detection of αSyn aggregation using the 5G4 antibody, with quantitative analysis confirming significant changes (n = 4). ( P ) Analysis of αSyn partitioning in soluble and insoluble brain fractions by western blot, with quantitative data indicating significant alterations (n = 4). Data are expressed as means ± SEM, * P < 0.05, ** P < 0.01, *** P < 0.001, ns, not significant.

Article Snippet: Soluble and insoluble protein fractions were isolated using a commercially accessible Soluble & Insoluble Protein Extraction Kit (Sangon Biotech, Shanghai, China).

Techniques: Olfactory, Modification, Activity Assay, Immunostaining, Western Blot, Expressing, Staining