Journal: Frontiers in Immunology

Article Title: Mucosal-Associated Invariant T Cells Improve Nonalcoholic Fatty Liver Disease Through Regulating Macrophage Polarization

doi: 10.3389/fimmu.2018.01994

Figure Lengend Snippet: Comparisons of MAIT cell distribution and MR1 expression in livers between patients and HC. (A) Representative confocal staining of CD3 (in green), MR1-5-OP-RU TEM (in red) and DAPI (for nuclei in blue) in the livers of HC, and patients with NAS ≤ 4 ( n = 22), NAS ≥ 5 ( n = 18) (original magnification, × 400). (B) The number of hepatic MAIT cells in NAFLD patients increased compared with HC and was positively correlated with NAS in NAFLD patients. (C) Representative immunohistochemical staining of MR1 in liver of HC and NAFLD patients (original magnification, × 200). (D) Representative confocal staining of CD68 + (in green), MR1 (in red) and DAPI (for nuclei in blue) (original magnification, × 400). (E) Representative overlaid histogram plots of MR1 staining of CD14 + monocytes-derived macrophages with FFA stimulation and BSA as control (Left panel). Statistical analysis of MFI of MR1 staining depicted an elevated level of MR1 expression on macrophages after FFA stimulation (Right panel) ( n = 5). Data were analyzed with Mann–Whitney U- test. * P

Article Snippet: After quenching endogenous peroxidase activity with 3% H2 O2 in methanol, the sections were incubated in 5% bovine serum albumin (BSA) followed by incubation with antibodies against MR1 (1:300, monoclone 26.5, Santa Cruz) for immunohistochemistry, CD3 (1:200) and MR1 tetramer [TEM, 1:500, provided by the National Institutes of Health (NIH) tetramer facility] for immunofluorescence overnight at 4°C.

Techniques: Expressing, Staining, Transmission Electron Microscopy, Immunohistochemistry, Derivative Assay, MANN-WHITNEY

Journal: PLoS ONE

Article Title: The S100A10 Subunit of the Annexin A2 Heterotetramer Facilitates L2-Mediated Human Papillomavirus Infection

doi: 10.1371/journal.pone.0043519

Figure Lengend Snippet: HPV16 PsV infection decreases following SLPI treatment or anti-annexin A2 antibody inhibition of A2t. HaCaT cells were infected with HPV16 pseudovirions containing a GFP plasmid. Infectivity was scored at 48 h post infection by enumerating GFP-positive cells by flow cytometry. (A) Cells were preincubated with increasing amounts of SLPI or BSA for one hour at 4° prior to PsV infection. The mean percentage of HPV16 PsV infected cells (GFP-positive) normalized to the PsV only group ± SD are presented. (** P

Article Snippet: For SLPI blocking experiments, the cells were incubated with increasing amounts of rhu SLPI or BSA (Bio-Rad, Hercules, CA) in PBS for 1 h at 4°C prior to addition of PsV.

Techniques: Infection, Inhibition, Plasmid Preparation, Flow Cytometry, Cytometry

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Propionate-induced changes in cardiac metabolism, notably CoA trapping, are not altered by l-carnitine

doi: 10.1152/ajpendo.00081.2018

Figure Lengend Snippet: Short-chain dicarboxylylcarnitines have little 13 C labeling in the perfused hearts with [ 13 C 6 ]glucose and [1- 13 C]palmitate. Rat hearts were perfused with 40% 11 mM [ 13 C 6 ]glucose, [1- 13 C]palmitate, 0.1 mM 2-DG, 3% BSA, 100 µU insulin, and 50 µM

Article Snippet: The perfusions were conducted as the following groups: 1 ) hearts perfused with 11 mM 40% [13 C6 ]glucose, 0.4 mM [1-13 C]palmitate, 3% BSA (fatty acid free; Fisher Scientific), 100 µU/ml insulin, 50 µM l -carnitine, and 0.1 mM 2-DG ± (3 mM propionate ± 3 mM l -carnitine); 2 ) hearts perfused with 11 mM glucose, 0.4 mM palmitate, 3% BSA, 100 µU/ml insulin, 50 µM l -carnitine, and 3 mM [13 C3 ]propionate; 3 ) hearts perfused with 11 mM glucose, 100 µU/ml insulin, 50 µM l -carnitine, 3% BSA, and 0.2 mM [1-13 C]octanoate ± 3 mM propionate.

Techniques: Labeling

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Propionate-induced changes in cardiac metabolism, notably CoA trapping, are not altered by l-carnitine

doi: 10.1152/ajpendo.00081.2018

Figure Lengend Snippet: Propionate-mediated changes of acyl-CoAs and acylcarnitines in the perfused hearts. Rat hearts were perfused with 40% 11 mM [ 13 C 6 ]glucose, [1- 13 C]palmitate, 0.1 mM 2-DG, 3% BSA, 100 µU insulin, and 50 µM l -carnitine (control, n = 5), control

Article Snippet: The perfusions were conducted as the following groups: 1 ) hearts perfused with 11 mM 40% [13 C6 ]glucose, 0.4 mM [1-13 C]palmitate, 3% BSA (fatty acid free; Fisher Scientific), 100 µU/ml insulin, 50 µM l -carnitine, and 0.1 mM 2-DG ± (3 mM propionate ± 3 mM l -carnitine); 2 ) hearts perfused with 11 mM glucose, 0.4 mM palmitate, 3% BSA, 100 µU/ml insulin, 50 µM l -carnitine, and 3 mM [13 C3 ]propionate; 3 ) hearts perfused with 11 mM glucose, 100 µU/ml insulin, 50 µM l -carnitine, 3% BSA, and 0.2 mM [1-13 C]octanoate ± 3 mM propionate.

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Propionate-induced changes in cardiac metabolism, notably CoA trapping, are not altered by l-carnitine

doi: 10.1152/ajpendo.00081.2018

Figure Lengend Snippet: M3 acylcarnitine synthesis kinetics in the heart perfused with [ 2 H 3 ] l -carnitine. Rat hearts were perfused with 11 mM glucose, palmitate, 3% BSA, 100 µU insulin, and 50 µM [ 2 H 3 ] l -carnitine for 0, 2, 5, 10, 20, 40, and 60 min. There was

Article Snippet: The perfusions were conducted as the following groups: 1 ) hearts perfused with 11 mM 40% [13 C6 ]glucose, 0.4 mM [1-13 C]palmitate, 3% BSA (fatty acid free; Fisher Scientific), 100 µU/ml insulin, 50 µM l -carnitine, and 0.1 mM 2-DG ± (3 mM propionate ± 3 mM l -carnitine); 2 ) hearts perfused with 11 mM glucose, 0.4 mM palmitate, 3% BSA, 100 µU/ml insulin, 50 µM l -carnitine, and 3 mM [13 C3 ]propionate; 3 ) hearts perfused with 11 mM glucose, 100 µU/ml insulin, 50 µM l -carnitine, 3% BSA, and 0.2 mM [1-13 C]octanoate ± 3 mM propionate.

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Propionate-induced changes in cardiac metabolism, notably CoA trapping, are not altered by l-carnitine

doi: 10.1152/ajpendo.00081.2018

Figure Lengend Snippet: The labeling of metabolites from [ 13 C 3 ]propionate in the perfused hearts and livers. Rat hearts were perfused with 11 mM glucose, palmitate, 3% BSA, 100 µU insulin, 3 mM [ 13 C 3 ]propionate, and 50 µM l -carnitine ( n = 6). Liver perfusion

Article Snippet: The perfusions were conducted as the following groups: 1 ) hearts perfused with 11 mM 40% [13 C6 ]glucose, 0.4 mM [1-13 C]palmitate, 3% BSA (fatty acid free; Fisher Scientific), 100 µU/ml insulin, 50 µM l -carnitine, and 0.1 mM 2-DG ± (3 mM propionate ± 3 mM l -carnitine); 2 ) hearts perfused with 11 mM glucose, 0.4 mM palmitate, 3% BSA, 100 µU/ml insulin, 50 µM l -carnitine, and 3 mM [13 C3 ]propionate; 3 ) hearts perfused with 11 mM glucose, 100 µU/ml insulin, 50 µM l -carnitine, 3% BSA, and 0.2 mM [1-13 C]octanoate ± 3 mM propionate.

Techniques: Labeling

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Propionate-induced changes in cardiac metabolism, notably CoA trapping, are not altered by l-carnitine

doi: 10.1152/ajpendo.00081.2018

Figure Lengend Snippet: Short-chain dicarboxylylcarnitines are more labeled from [ 13 C 3 ]propionate in liver than in heart. Rat hearts were perfused with 11 mM glucose, palmitate, 3% BSA, 100 µU insulin, 50 µM l -carnitine, and 3 mM [ 13 C 3 ]propionate for 25 min.

Article Snippet: The perfusions were conducted as the following groups: 1 ) hearts perfused with 11 mM 40% [13 C6 ]glucose, 0.4 mM [1-13 C]palmitate, 3% BSA (fatty acid free; Fisher Scientific), 100 µU/ml insulin, 50 µM l -carnitine, and 0.1 mM 2-DG ± (3 mM propionate ± 3 mM l -carnitine); 2 ) hearts perfused with 11 mM glucose, 0.4 mM palmitate, 3% BSA, 100 µU/ml insulin, 50 µM l -carnitine, and 3 mM [13 C3 ]propionate; 3 ) hearts perfused with 11 mM glucose, 100 µU/ml insulin, 50 µM l -carnitine, 3% BSA, and 0.2 mM [1-13 C]octanoate ± 3 mM propionate.

Techniques: Labeling

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Propionate-induced changes in cardiac metabolism, notably CoA trapping, are not altered by l-carnitine

doi: 10.1152/ajpendo.00081.2018

Figure Lengend Snippet: Metabolites altered by propionate. Rat hearts were perfused with 40% 11 mM [ 13 C 6 ]glucose, [1- 13 C]palmitate, 0.1 mM 2-DG, 3% BSA, 100 µU insulin, and 50 µM l -carnitine (control, n = 5), control + 3 mM propionate (PA, n = 5), and PA + 3

Article Snippet: The perfusions were conducted as the following groups: 1 ) hearts perfused with 11 mM 40% [13 C6 ]glucose, 0.4 mM [1-13 C]palmitate, 3% BSA (fatty acid free; Fisher Scientific), 100 µU/ml insulin, 50 µM l -carnitine, and 0.1 mM 2-DG ± (3 mM propionate ± 3 mM l -carnitine); 2 ) hearts perfused with 11 mM glucose, 0.4 mM palmitate, 3% BSA, 100 µU/ml insulin, 50 µM l -carnitine, and 3 mM [13 C3 ]propionate; 3 ) hearts perfused with 11 mM glucose, 100 µU/ml insulin, 50 µM l -carnitine, 3% BSA, and 0.2 mM [1-13 C]octanoate ± 3 mM propionate.

Techniques:

Journal: American Journal of Physiology - Endocrinology and Metabolism

Article Title: Propionate-induced changes in cardiac metabolism, notably CoA trapping, are not altered by l-carnitine

doi: 10.1152/ajpendo.00081.2018

Figure Lengend Snippet: Propionate suppressed mitochondrial metabolism of octanoate to TCA. Rat hearts were perfused in Langendorff mode with 11 mM glucose, 0.2 mM [1- 13 C]octanoate, 3% BSA, 100 µU insulin, and 50 µM l -carnitine (M1 OA, n = 4), control + 3 mM

Article Snippet: The perfusions were conducted as the following groups: 1 ) hearts perfused with 11 mM 40% [13 C6 ]glucose, 0.4 mM [1-13 C]palmitate, 3% BSA (fatty acid free; Fisher Scientific), 100 µU/ml insulin, 50 µM l -carnitine, and 0.1 mM 2-DG ± (3 mM propionate ± 3 mM l -carnitine); 2 ) hearts perfused with 11 mM glucose, 0.4 mM palmitate, 3% BSA, 100 µU/ml insulin, 50 µM l -carnitine, and 3 mM [13 C3 ]propionate; 3 ) hearts perfused with 11 mM glucose, 100 µU/ml insulin, 50 µM l -carnitine, 3% BSA, and 0.2 mM [1-13 C]octanoate ± 3 mM propionate.

Techniques:

Journal: Frontiers in Oncology

Article Title: Recombinant heat shock protein 70 in combination with radiotherapy as a source of tumor antigens to improve dendritic cell immunotherapy

doi: 10.3389/fonc.2012.00149

Figure Lengend Snippet: Exogenous rHsp70 did not increase radiation–induced CT26/PSA apoptosis nor increase BM-DC phagocytosis. (A) The percentage of apoptotic CT26/PSA cells was analyzed by annexin-V and PI (propidium iodide) staining (shown in the upper right corner) in, left to right, parental CT26/PSA cells in PBS (parental CT26) or irradiated CT26/PSA cells (irradiated CT26) in PBS (PBS) or 10 μg/ml of BSA (BSA), rHsp70 (HSP70), or rHsp70C′-PSA (Hsp70/PSA) for 24 h incubation. The number of percentage was indicated AnnexinV-positive cells. (B) Cell Tracker-labeled CT26/PSA (5 × 10 5 ), pretreated as in Figure 1A , and PKH-67-labeled BM-DCs (5 × 10 5 ) were cocultured in 6-well plates for 24 h at 4°C (top row, no phagocytosis control) or 37°C (bottom row) and the percentages of double-positive cells measured. The panels are arranged in the same order as in Figure 1A . (C) Untreated (lane C) or irradiated CT26/PSA cells (lane RT) or control cells heated at 42°C for 30 min (positive control; lane 42) were harvested for Western blot analysis for Hsp70 protein in the cell lysate and culture supernatant. Lane rHsp is rHsp70. These data are representative of those obtained in three experiments.

Article Snippet: The BM-DCs (5−10 × 105 ) were stained with 50 μl of FITC-conjugated antibodies in phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA) and 0.1% azide, which also served as the washing buffer, then were subjected to fluorescence-activated cell sorting (FACS) analysis using a FASCalibur flow cytometer (BD Bioscience, San Diego, CA, USA).

Techniques: Staining, Irradiation, Incubation, Labeling, Positive Control, Western Blot

Journal: Frontiers in Oncology

Article Title: Recombinant heat shock protein 70 in combination with radiotherapy as a source of tumor antigens to improve dendritic cell immunotherapy

doi: 10.3389/fonc.2012.00149

Figure Lengend Snippet: Exogenous rHSPs enhance the expression of pro-inflammatory cytokines. BM-DCs (1.5 × 10 6 in 1 ml) were incubated with medium alone (control) or LPS (10 μg/ml) or were cocultured at 37°C for 24 h with irradiated-CT26/PSA cells (1.5 × 10 6 in 1 ml) of PBS or 10 μg/ml of BSA, rHsp70, or rHsp70C′-PSA in the absence or presence of 10 μg/ml of polymyxin B. The supernatants were then harvested and the concentration of IL-12 (A) or TNF-α (B) measured by ELISA. The data are the mean ± SD for three-independent experiments. * Indicates p

Article Snippet: The BM-DCs (5−10 × 105 ) were stained with 50 μl of FITC-conjugated antibodies in phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA) and 0.1% azide, which also served as the washing buffer, then were subjected to fluorescence-activated cell sorting (FACS) analysis using a FASCalibur flow cytometer (BD Bioscience, San Diego, CA, USA).

Techniques: Expressing, Incubation, Irradiation, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Oncology

Article Title: Recombinant heat shock protein 70 in combination with radiotherapy as a source of tumor antigens to improve dendritic cell immunotherapy

doi: 10.3389/fonc.2012.00149

Figure Lengend Snippet: Irradiated-CT26/PSA cells cause BM-DC maturation in the presence of rHSPs. BM-DCs (1.5 × 10 6 in 1 ml) were incubated with medium alone (control) or LPS (10 μg/ml) or were cocultured with irradiated-CT26/PSA cells (1.5 × 10 6 in 1 ml) in PBS or 10 μg/ml of BSA, rHsp70, or rHsp70C′-PSA. After 48 h, the cells were harvested and stained with antibodies against MHC class II molecule I-A d and LAMP1. Column 1, DAPI nuclear staining; column 2, LAMP1 staining; column 3, MHC class II intracellular staining; column 4, merged images. Images were acquired using confocal microscopy at a magnification of ×400. This experiment was repeated three times with reproducible results. Scale bars, 20 μm.

Article Snippet: The BM-DCs (5−10 × 105 ) were stained with 50 μl of FITC-conjugated antibodies in phosphate-buffered saline (PBS) containing 1% bovine serum albumin (BSA) and 0.1% azide, which also served as the washing buffer, then were subjected to fluorescence-activated cell sorting (FACS) analysis using a FASCalibur flow cytometer (BD Bioscience, San Diego, CA, USA).

Techniques: Irradiation, Incubation, Staining, Confocal Microscopy