Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Protease Domain in HEV pORF1 Mediates the Replicase’s Localization to Multivesicular Bodies and Its Exosomal Release

doi: 10.1016/j.jcmgh.2024.01.001

Figure Lengend Snippet: HEV pORF1 predominantly localizes to CD63-positive vesicular structures. Analyses of subcellular distribution of pORF1 ectopically expressed in A549/D3 cells. ( A ) Confocal microscopy of double-tagged pORF1; EGFP-mCherry Merge displays the subcellular distribution of EGFP ( green )-mCherry ( red ) fusion protein without insertion of pORF1. C-term, C-terminal mCherry; N-term, N-terminal EGFP. ( B ) Particle analysis of only C-terminus–containing vesicles (C-term) and N- and C-terminus-containing vesicles (C+N-terminus). ∗∗∗ P > .0001; ∼300 particles were analyzed. ( C ) Confocal microscopy of C-terminally mScarlet-tagged pORF1 ( red ) and indirect immunofluorescent stain of different organelle marker proteins ( green ); ERGIC53, ERGIC; GM130, Golgi; EEA1, EEs; Rab11a, REs; LC3B, APs; LAMP2, LYs; CD63, MVBs. ( D ) Particle analysis of pORF1-containing vesicles (pORF1 alone) and pORF1- and marker protein–containing vesicles (pORF1 + marker); between ∼200 and ∼1000 particles were analyzed for each organelle marker. Single channels depicted as greyscale images with headers indicating the channel coloring; zoom refers to areas magnified from white rectangles in merge; white ellipses indicate nucleus; scale bar represents 40 μm. APs, autophagosomes; EEs, early endosomes; ERGIC, ER-Golgi intermediate compartment; Golgi, Golgi apparatus; LYs, lysosomes; MVBs, multivesicular bodies; REs, recycling endosomes. Data are displayed as mean ± standard error of the mean. Microscopy performed on Leica TCS SP8 System with 100× objective (numerical arperture 1.4). Images deconvoluted via lightning algorithm using LAS X Control software.

Article Snippet: Primary antibodies were raised against ERGIC53/LMAN1 (lectin mannose-binding 1; Santa Cruz Biotechnologies, sc-365158), GM130/GOLGA2 (golgin A2; Cell Signaling, 12480S), EEA1 (early endosome antigen 1; Abcam, ab206860), Rab11a (Ras-related protein Rab-11A; ProSci, 33-825), (MAP1)LC3B (microtubule-associated protein 1 light chain 3-beta; MBL International, PM036 MBL), LAMP2/CD107b (lysosomal associated membrane protein 2; BD Biosciences, 555803), and CD63 (cluster of differentiation 63; Abcam, ab59479).

Techniques: Confocal Microscopy, Particle Size Analysis, Staining, Marker, Microscopy, Control, Software

Journal: EBioMedicine

Article Title: Rapid diagnostics for point-of-care quantification of soluble transferrin receptor

doi: 10.1016/j.ebiom.2019.03.017

Figure Lengend Snippet: (A) Representative images of the test and control lines on the sTfR test strip for a range of sTfR concentrations in spiked-buffer standards (B) Calibration curve with T/C values for a range of sTfR concentrations in spiked-buffer standards.

Article Snippet: Results demonstrated a correlation of 0·93 (P < 0.0001), with an R 2 value of 0·87 with linear fitting applied on the trendline. (A) Representative images of the test and control lines on the sTfR test strip for a range of concentrations of serum samples (B) Correlation plot of predicted serum sTfR concentrations based on comparison of point-of-care test results with Ramco ELISA (C) Results of bootstrapping to compute the resulting correlation coefficients.

Techniques: Stripping Membranes

Journal: EBioMedicine

Article Title: Rapid diagnostics for point-of-care quantification of soluble transferrin receptor

doi: 10.1016/j.ebiom.2019.03.017

Figure Lengend Snippet: (A) Representative images of the test and control lines on the sTfR test strip for a range of sTfR concentrations of in ‘Access’ calibrators (B) Calibration curve with T/C values for a range of sTfR concentrations of in ‘Access’ calibrators.

Article Snippet: Results demonstrated a correlation of 0·93 (P < 0.0001), with an R 2 value of 0·87 with linear fitting applied on the trendline. (A) Representative images of the test and control lines on the sTfR test strip for a range of concentrations of serum samples (B) Correlation plot of predicted serum sTfR concentrations based on comparison of point-of-care test results with Ramco ELISA (C) Results of bootstrapping to compute the resulting correlation coefficients.

Techniques: Stripping Membranes

Journal: EBioMedicine

Article Title: Rapid diagnostics for point-of-care quantification of soluble transferrin receptor

doi: 10.1016/j.ebiom.2019.03.017

Figure Lengend Snippet: (A) Representative images of the test and control lines on the sTfR test strip for a range of concentrations of serum samples (B) Correlation plot of predicted serum sTfR concentrations based on comparison of point-of-care test results with Ramco ELISA (C) Results of bootstrapping to compute the resulting correlation coefficients.

Article Snippet: Results demonstrated a correlation of 0·93 (P < 0.0001), with an R 2 value of 0·87 with linear fitting applied on the trendline. (A) Representative images of the test and control lines on the sTfR test strip for a range of concentrations of serum samples (B) Correlation plot of predicted serum sTfR concentrations based on comparison of point-of-care test results with Ramco ELISA (C) Results of bootstrapping to compute the resulting correlation coefficients.

Techniques: Stripping Membranes, Enzyme-linked Immunosorbent Assay