Journal: Cells

Article Title: Empagliflozin Preserves Cardiomyocyte Structural Homeostasis via the Stabilization of the Integrin α5–Desmocollin-2 Adhesion Axis in Sepsis-Induced Cardiomyopathy

doi: 10.3390/cells14181452

Figure Lengend Snippet: Empagliflozin ameliorates LPS-induced cardiac dysfunction and myocardial injury. ( A ) Echocardiography panels: top, B-mode long-axis image; middle, M-mode from the parasternal short-axis at the papillary muscle level; bottom, pulsed-wave Doppler of transmitral inflow (E and A waves). ( B ) Quantification of cardiac function parameters: Ejection Fraction (EF), Fractional Shortening (FS), and E/A Ratio. ( C ) Representative images of Hematoxylin and Eosin (H & E) staining of myocardial tissue sections showing inflammatory infiltration. Longitudinal and cross sections are shown with a scale bar of 1 mm. Magnified views of perivascular and interstitial regions are shown with a scale bar of 20 µm; green arrows indicate inflammatory cells. ( D ) Representative images of Masson’s trichrome staining revealing myocardial fibrosis. Longitudinal sections are shown with a scale bar of 1 mm, and cross sections with a scale bar of 0.5 mm. Magnified views of perivascular and interstitial regions are shown with a scale bar of 40 µm; yellow arrows indicate collagen deposition (fibrotic areas). ( E ) Quantification of the fibrotic area from Masson’s trichrome staining. ( F ) Serum concentrations of cardiac troponin T (cTnT) and C-reactive protein (CRP) measured by ELISA. Data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA. **** p < 0.0001, *** p < 0.001, ** p < 0.01.

Article Snippet: Serum levels of cardiac troponin T (cTnT) and C-reactive protein (CRP) were quantified using commercial enzyme-linked immunosorbent assay (ELISA) kits (cTnT: Cat. No. orb565454, Biorbyt, Wuhan, China; CRP: Cat. No. orb219654, Biorbyt, Wuhan, China) according to the manufacturer’s instructions.

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Cell

Article Title: Two-component spike nanoparticle vaccine protects macaques from SARS-CoV-2 infection

doi: 10.1016/j.cell.2021.01.035

Figure Lengend Snippet:

Article Snippet: Five-fold serial dilutions of polyclonal macaque (Molecular Innovations) or human IgG, starting at a concentration of 1 μg/mL, were added to wells containing the coated goat anti-Human IgG λ and κ.

Techniques: Purification, Recombinant, Mass Spectrometry, Sequencing, Ligation, Luciferase, Enzyme-linked Immunospot, Plasmid Preparation, Software, Chromatography, Luminex, Expressing

Journal: JCI Insight

Article Title: An effective cell-penetrating antibody delivery platform

doi: 10.1172/jci.insight.127474

Figure Lengend Snippet: (A) Tumor growth kinetics when treated with indicated modified antibodies (n = 8). SD shown. Two-way ANOVA (Tukey’s multiple-comparisons test) was used: ***P < 0.001. (B) ELISA on pY(418)-Src recognition by anti-pY(418)Src antibody upon modification performed twice in triplicates. (C) Representative Western blot assessing gene expression in tissue homogenates isolated from tumors. (D) Tumor tissue integrity, tumor accumulation of modified antibodies and CD31+ tumor vasculature, and pY(418)-Src levels shown by confocal microscopy (left) with quantification (right). (E) Representative RT-PCR on Src kinase target gene mRNA expression in tumors treated as indicated, performed in triplicates. (B, D, and E) SD shown. Unpaired Student’s t test: **P < 0.01, and ***P < 0.001.

Article Snippet: Human IgG λ chain was detected using the human IgG λ light chain ELISA kit according to the manufacturer’s instructions (MyBioSource.com).

Techniques: Modification, Enzyme-linked Immunosorbent Assay, Western Blot, Gene Expression, Isolation, Confocal Microscopy, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Scientific Reports

Article Title: Immunoglobulin G Expression in Human Sperm and Possible Functional Significance

doi: 10.1038/srep20166

Figure Lengend Snippet: ( a ) The primary antibody is rabbit anti-human IgG γ chain, the secondary antibody is donkey anti-rabbit IgG (H + L) labeled with Alexa Fluor® 594 and the positive stain is red in color. ( b ) The primary antibody is mouse anti-human IgG κ chain antibody, the secondary antibody is goat anti-mouse labeled with Alexa Fluor® 488 and the positive stain is green in color. ( c ) The primary antibody is rabbit anti-human IgG λ chain, and the secondary antibody is donkey anti-rabbit Alexa Fluor® 594 IgG (H + L) and the positive stain is red in color. ( d ) Negative control. DAPI was used to stain the nucleus of sperm (blue). The scale bar is 30 μm for all photos.

Article Snippet: The prepared motile sperms were incubated at 37 °C in a CO 2 (5%) incubator for 4 h in a 100 μl drop of capacitated liquid, containing 1 μl of primary antibody such as rabbit anti-human IgG γ chain polyclonal antibody (1:100, Dako, Denmark), rabbit anti-human IgG λchain monoclonal antibody (1:100, ZSGB-BIO, China) or mouse anti-human IgG κ chain polyclonal antibody (1:100, Dako, Denmark) (See detail in ).

Techniques: Labeling, Staining, Negative Control

Journal: Scientific Reports

Article Title: Immunoglobulin G Expression in Human Sperm and Possible Functional Significance

doi: 10.1038/srep20166

Figure Lengend Snippet: Normal human sperm was processed and stained with rabbit anti-human IgG antibody (γ chain specific) and the secondary antibody to rabbit IgG was labeled with 10 nm colloidal gold. The positive stain was shown by highly electron-dense gold particles. ( a ) The neck region of sperm contains abundant gold particles (white arrow) showing the subcellular distribution of IgG. Nu is nucleus and Ce is centriole. ( b ) Lower magnification of ( a ). ( c ) The middle piece of a sperm tail. The gold particles (white arrow) depict IgG distribution. Mi is mitochondria and Ax is axoneme. ( d ) Lower magnification of ( c ). The results demonstrate that both the neck region and the middle piece of the tail of human sperm contain synthesis IgG γ chain molecule.

Article Snippet: The prepared motile sperms were incubated at 37 °C in a CO 2 (5%) incubator for 4 h in a 100 μl drop of capacitated liquid, containing 1 μl of primary antibody such as rabbit anti-human IgG γ chain polyclonal antibody (1:100, Dako, Denmark), rabbit anti-human IgG λchain monoclonal antibody (1:100, ZSGB-BIO, China) or mouse anti-human IgG κ chain polyclonal antibody (1:100, Dako, Denmark) (See detail in ).

Techniques: Staining, Labeling

Journal: Scientific Reports

Article Title: Immunoglobulin G Expression in Human Sperm and Possible Functional Significance

doi: 10.1038/srep20166

Figure Lengend Snippet: ( a ) Results of Western blot. The first lane is total protein extracted from human sperm (56 μg), the second lane to the forth lane is 0.2 μg /0.5 μg /2 μg human standard IgG molecule respectively (positive control). Anti-human IgG γ chain antibody, anti-IgG λ chain antibody and anti-IgG κ chain antibody were used as primary antibodies respectively. Molecular weight of protein was labeled on the right of each panel. ( b ) Expression of RAG-1 (327 bp) and RAG-2 (193 bp) genes were detected in human spermatozoa. Lane 1 is PCR product of RAG1 and RAG2 gene extracted from Raji cells (positive control). Lane 2 is DNase treated cDNA of Raji cells was used as a template (negative control). Lane 3 is cDNA of normal human sperm extract was used as a template. RAG1 and RAG2 genes were amplified. Lane 4 is DNase treated template of normal human sperm. No RAG1 or RAG2 gene was detected. ( c ) RT-PCR amplification of mRNA transcripts of IgG λ chain, AID, IGHG1, CD19 and GAPDH in human sperm. There was no contamination from B cells in human sperm since CD19 was negative. Human sperm could express transcripts of IgG λ chain, AID, IGHG1 and GAPDH. M is DNA ladder, W is water as a negative control, S represents human sperm, Raji cell was used as the positive control. CD19 is a marker for Raji cell. The results show that human sperm can express the essential genes for immunoglobulin G V-D-J rearrangement. In addition, human sperm can synthesis IgG in both the protein level and the mRNA levels.

Article Snippet: The prepared motile sperms were incubated at 37 °C in a CO 2 (5%) incubator for 4 h in a 100 μl drop of capacitated liquid, containing 1 μl of primary antibody such as rabbit anti-human IgG γ chain polyclonal antibody (1:100, Dako, Denmark), rabbit anti-human IgG λchain monoclonal antibody (1:100, ZSGB-BIO, China) or mouse anti-human IgG κ chain polyclonal antibody (1:100, Dako, Denmark) (See detail in ).

Techniques: Western Blot, Positive Control, Molecular Weight, Labeling, Expressing, Negative Control, Amplification, Reverse Transcription Polymerase Chain Reaction, Marker

Journal: Scientific Reports

Article Title: Immunoglobulin G Expression in Human Sperm and Possible Functional Significance

doi: 10.1038/srep20166

Figure Lengend Snippet: ( a ) In situ hybridization assay detected the expression and location of IgG mRNA. (A,B) were human sperms incubated with anti-sense probe and sense probe against IgG γ chain respectively. The positive stain (purple) in (A) demonstrated mRNA expression of IgG which was mainly located in the neck of human sperm. There was no positive stain with sense probe as shown in (B). (C,D) were human normal spleen tissue served as additional controls. (C) was incubated with anti-sense probe against IgG γ chain (positive control) and (D) was incubated with sense probe against IgG γ chain (negative control). Bar was 20 μm in (A,B). Bar was 25 μm in (C,D). ( b ) Anti-IgG antibody could inhibit sperm-egg penetration. After being pre-incubated with anti-human IgG antibody (γ chain specific) for 4 h at 37 °C, zona-free hamster egg-sperm penetration assay were performed and the penetration ratio was calculated. The sperms without anti-IgG antibody treatment were used as a control for comparison. Data show means ± SE of four independent experiments. The mean penetration ratio in the treatment group is 27%, while the mean ratio in the control group is 15%. The difference was statistically significant between the two groups (*p < 0.05, One way ANOVA, Dunnet’s post-hoc test). n is the number of zona-free hamster eggs used in the penetration assay for each group. The result shows that IgG produced by sperm may play a role in sperm-egg fusion.

Article Snippet: The prepared motile sperms were incubated at 37 °C in a CO 2 (5%) incubator for 4 h in a 100 μl drop of capacitated liquid, containing 1 μl of primary antibody such as rabbit anti-human IgG γ chain polyclonal antibody (1:100, Dako, Denmark), rabbit anti-human IgG λchain monoclonal antibody (1:100, ZSGB-BIO, China) or mouse anti-human IgG κ chain polyclonal antibody (1:100, Dako, Denmark) (See detail in ).

Techniques: In Situ Hybridization, Expressing, Incubation, Staining, Positive Control, Negative Control, Control, Comparison, Produced