Journal: Scientific Reports

Article Title: Intravenous high mobility group box 1 fragment improves cardiac function, fibrosis, and coronary flow in porcine ischemic cardiomyopathy model

doi: 10.1038/s41598-026-38592-7

Figure Lengend Snippet: RT-PCR analysis. Comparison of mRNA expression in the border and remote zones between the two groups using RT-PCR. (7A) SDF1, (7B) HGF, (7C) VEGFα, (7D) FGF2, (7E) TGFβ3, (7F) IL-10, (7G) PDGFRα, (7H) PDGFRβ. *p < 0.05, **p < 0.01.

Article Snippet: The following genes were analyzed using TaqMan Gene Expression Assay (Applied Biosystems): SDF1 (Ss03391855_m1), HGF (Ss06927387_m1), VEGFA (Ss03393990_m1), FGF2 (Ss03375809_u1), TGFβ3 (Ss03394351_m1), IL-10 (Ss03382372_u1), PDGFRα (Ss04955107_g1), and PDGFRβ (Ss06879561_m1).

Techniques: Reverse Transcription Polymerase Chain Reaction, Comparison, Expressing

Journal: bioRxiv

Article Title: Mechanism of giant magnetic field effect in fluorescence of mScarlet3, a red fluorescent protein

doi: 10.1101/2025.02.26.640351

Figure Lengend Snippet: E. coli expressing red-emitting fluorescent proteins were grown on LB (lysogeny broth) agar plates and then imaged in an epifluorescence microscope. Panels show responses to a 10 mT magnetic field (grey bars). A 595 nm longpass emission filter was used. A) mScarlet-I3, B) mScarlet3, C) mRuby3, D) mSandy2, E) mKate2.

Article Snippet: The additional fluorescent proteins tested were mScarlet-I3 (Addgene #189757), mRuby3 (Addgene #104005), mSandy2 (Addgene #177760), and mKate2 (Addgene #104030).

Techniques: Expressing, Microscopy

Journal: Journal of proteome research

Article Title: Integrated approaches for analyzing U1-70K cleavage in Alzheimer's disease.

doi: 10.1021/pr5003593

Figure Lengend Snippet: Figure 1. N40K occurrence is independent of the postmortem interval and correlates with the decrease of the U1-70K level in AD. (A) Identification of N40K in the detergent insoluble fraction of AD brain by 1D SDS gel and LC−MS/MS. Ctl: control case. (B) Validation of N40K only in AD (case 8 in Supporting Information Table S1) by Western blotting. (C) No statistically significant difference in postmortem intervals between the N40K negative and positive cases. (D) Western blotting to show reduced U1-70K level in the total homogenate in N40K positive cases. The numbers above the gel indicate the corresponding case numbers in Supporting Information Table S1. (E) Quantitation of the Western blotting intensity in panel D to indicate an inverse correlation of U1-70K in total homogenate with insoluble N40K. Relative intensities of U1-70K and N40K were normalized by setting the maximal values to 100, respectively.

Article Snippet: Fulllength U1-70K and its truncations were cloned from a commercial plasmid (U1-70K-myc-DDK, pCMV6, Origene, RC201713).

Techniques: SDS-Gel, Liquid Chromatography with Mass Spectroscopy, Control, Biomarker Discovery, Western Blot, Quantitation Assay

Journal: Journal of proteome research

Article Title: Integrated approaches for analyzing U1-70K cleavage in Alzheimer's disease.

doi: 10.1021/pr5003593

Figure Lengend Snippet: Figure 2. Identification of U1-70K and N40K peptides in AD. These peptides are shown in different colors in the protein sequence, together with peptides monitored in LC−SRM.

Article Snippet: Fulllength U1-70K and its truncations were cloned from a commercial plasmid (U1-70K-myc-DDK, pCMV6, Origene, RC201713).

Techniques: Sequencing

Journal: Journal of proteome research

Article Title: Integrated approaches for analyzing U1-70K cleavage in Alzheimer's disease.

doi: 10.1021/pr5003593

Figure Lengend Snippet: Figure 3. Analysis of the N40K C-terminus by LC−SRM. (A) Work flow of the method. (B) Identification and size confirmation of purified recombinant U1-70K by Western blotting, followed by band excision and in-gel digestion. (C) Identification of a N40K−containing gel band by Western blotting. The 10 kDa ladder was also used for precise alignment of immunoblotting images with stained gel. (D) The intensity ratios of light/heavy (L/H) of U1-70K peptide pairs, leading to the predicted cleavage region.

Article Snippet: Fulllength U1-70K and its truncations were cloned from a commercial plasmid (U1-70K-myc-DDK, pCMV6, Origene, RC201713).

Techniques: Recombinant, Western Blot, Staining

Journal: Journal of proteome research

Article Title: Integrated approaches for analyzing U1-70K cleavage in Alzheimer's disease.

doi: 10.1021/pr5003593

Figure Lengend Snippet: Figure 4. Analysis of the N40K N-terminus by MS. (A−C) MS/MS spectra for identifying the N-terminal peptide with methionine removal and threonine acetylation in U1-70K, N40K, and purified U1-70K in HEK293 cells, respectively. Matched ions were assigned by arrows indicating specific b or y ions.

Article Snippet: Fulllength U1-70K and its truncations were cloned from a commercial plasmid (U1-70K-myc-DDK, pCMV6, Origene, RC201713).

Techniques: Tandem Mass Spectroscopy

Journal: Journal of proteome research

Article Title: Integrated approaches for analyzing U1-70K cleavage in Alzheimer's disease.

doi: 10.1021/pr5003593

Figure Lengend Snippet: Figure 5. Determination of N40K C-terminus by Western blotting. (A) Expression of U1-70K N-terminal truncations in HEK293 cells. (B) Analysis of N40K in AD and three recombinant truncations. (C) Diagram of U1-70K domains and predicted N40K cleavage region. (D) N40K truncations covering all possible cleavage products with different C-terminal residues.

Article Snippet: Fulllength U1-70K and its truncations were cloned from a commercial plasmid (U1-70K-myc-DDK, pCMV6, Origene, RC201713).

Techniques: Western Blot, Expressing, Recombinant

Journal: Journal of proteome research

Article Title: Integrated approaches for analyzing U1-70K cleavage in Alzheimer's disease.

doi: 10.1021/pr5003593

Figure Lengend Snippet: Figure 6. Toxicity analysis of N40K in rat hippocampal primary neurons. (A) Restriction enzyme digestion at the cloning sites by BamHI and EcoRI to show equal levels of the plasmids of U1-70K and six N40K truncations. (B) Western blotting to show similar protein expression in HEK293 cells. (C) Co-transfection of nine plasmids each with RFP by the calcium phosphate method. (D) Quantitative analysis by counting degenerating neurons. Error bars represent standard errors of mean (SEM) of analyzed neurons in duplicated plates of the same batch of neuronal culture. The experiments were repeated three times with different batches of culture, all showing similar results. The asterisks indicate p < 0.05 by the ANOVA and Tukey range test. (E) The MTT assay of neuronal viability upon Lentiviral N40K transfection. Error bars represent SEM. The double asterisks indicate p < 0.01 by Student’s t-test.

Article Snippet: Fulllength U1-70K and its truncations were cloned from a commercial plasmid (U1-70K-myc-DDK, pCMV6, Origene, RC201713).

Techniques: Cloning, Western Blot, Expressing, Cotransfection, MTT Assay, Transfection