Journal: mBio

Article Title: Hepatitis C virus infects and perturbs liver stem cells

doi: 10.1128/mbio.01318-23

Figure Lengend Snippet: Liver organoids grow from HCV-infected individuals and show similar differentiation potential. ( A ) Representative brightfield microscopy images of liver organoids grown from uninfected (NV) or HCV + donors are shown in the stem cell (EM) and differentiated (DM) states. Organoids are morphologically distinct in EM vs DM states, but each state was morphologically identical across all six NV and HCV + donors. ( B ) Quantitative PCR (qPCR) quantification of hepatocyte stem cell marker LGR5 and hepatocyte markers ALB, CYP3A4, and CYP2B6 in DM organoids from three NV donors and three HCV + donors, and in two primary hepatocyte samples relative to EM. For each gene, data were pooled from n ≥ 2 biological replicates per organoid or hepatocyte donor and represented as mean ± SD. Transcript expression was normalized to 18S and plotted as a fold change over the gene’s expression in EM (ΔΔC T ) which was set to one and is marked by a dotted line. Fold change was plotted on a log10 scale. ( C ) qPCR quantification of HCV entry markers, CD81, OCLN, CLDN1, and SR-B1 in the same samples as in ( B ). EM expression levels were set to one (marked by a dotted line), and fold change in DM was plotted on a linear scale. ( D ) Representative light-sheet microscopy images are shown for differentiated liver organoids (DM) stained for hepatocyte markers HNF4α and ALB, HCV entry factors CLDN1 and CD81, or apical membrane marker ZO1.

Article Snippet: Primary antibodies we used include CD81 (BD Pharmigen, JS-81), claudin-1 (Thermo Scientific, 2H10D10), HNF4α (Cell Signaling Technology, C11F12), albumin (Sigma-Aldrich, HSA-11), and ZO1 (Thermo Scientific, 1A12).

Techniques: Infection, Microscopy, Real-time Polymerase Chain Reaction, Marker, Expressing, Staining, Membrane

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Delivery of mesenchymal stem cells-derived extracellular vesicles with enriched miR-185 inhibits progression of OPMD.

doi: 10.1080/21691401.2019.1623232

Figure Lengend Snippet: Figure 1. Characterization of miR-185 enriched MSC-EVs. (A) Isolated miR-185 MSC-EVs were characterized by TEM. Representative image is shown (scale bar ¼ 50 nm). (B) NTA was performed on the EVs to determine their concentration and size. EV diameter was measured and represented as mean ± SD, n ¼ 3 independent experiments performed in triplicate. Western blotting (insets) for EV markers CD9, CD81 and flotillin-1. (C) qPCR analysis for expression of miR-185 in EVs. n ¼ 5 inde- pendent experiments performed in triplicate (p < .01 versus Ctrl).

Article Snippet: MSC-EVs were probed with primary antibodies against CD81 (1:400 dilution, ProSci, Collins, CO), CD9 (1:300 dilution, Lifespan Bioscience Inc., Seattle, WA) and flotillin-1 (1:500 dilution, Abcam, Cambridge, MA) by Western blotting.

Techniques: Isolation, Concentration Assay, Western Blot, Expressing

Journal: American Journal of Cancer Research

Article Title: Circ_0006790 carried by bone marrow mesenchymal stem cell-derived exosomes regulates S100A11 DNA methylation through binding to CBX7 in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: Exo repress immune escape and immunosuppression in PDAC. (A) The protein expression of PD-L1 and CTLA-4 in PDAC cells treated with PBS or Exo. (B) Co-culture Exo- or PBS-treated PDAC cells with activated T cells. (C) Surviving PDAC cells examined using crystal violet staining. (D) The levels of IFN-γ and TNF-α, the immune effectors released by T cells, examined by ELISA. (E) Volume changes of xenograft tumors. (F) Weight of xenograft tumors. (G) Immunohistochemical detection of CD8+ T cell infiltration and expression of PD-L1 and CTLA-4 in xenograft tumors. *P < 0.05. Data are shown as mean ± SD of three technical replicates (n = 5). Unpaired t-test (F) and two-way ANOVA (A, C-E, G) were used for data comparison.

Article Snippet: The sections were sealed with 5% BSA at 37°C for 30 min and incubated overnight at 4°C with primary antibodies to CD8 (1:2000, ab209775, Abcam), PD-L1 (1:1.000, ab237726, Abcam), and CTLA-4 (1:500, ab237712, Abcam).

Techniques: Expressing, Co-Culture Assay, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining

Journal: American Journal of Cancer Research

Article Title: Circ_0006790 carried by bone marrow mesenchymal stem cell-derived exosomes regulates S100A11 DNA methylation through binding to CBX7 in pancreatic ductal adenocarcinoma

doi:

Figure Lengend Snippet: The circ_6790/CBX7/S100A11 axis regulates immune escape of PDAC cells. (A) The protein expression of PD-L1 and CTLA-4 in PDAC cells. (B) Surviving PDAC cells examined using crystal violet staining. (C) The levels of IFN-γ and TNF-α, the immune effectors released by T cells, examined by ELISA. (D) Volume changes of xenograft tumors. (E) Weight of xenograft tumors. (F) Immunohistochemical detection of CD8+ T cell infiltration and expression of PD-L1 and CTLA-4 in xenograft tumors. *P < 0.05. Data are shown as mean ± SD of three technical replicates (n = 5). One-way (B, E) or two-way ANOVA (A, C, D, F) were used for data comparison.

Article Snippet: The sections were sealed with 5% BSA at 37°C for 30 min and incubated overnight at 4°C with primary antibodies to CD8 (1:2000, ab209775, Abcam), PD-L1 (1:1.000, ab237726, Abcam), and CTLA-4 (1:500, ab237712, Abcam).

Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining