Journal: Viruses

Article Title: Recombinant Pseudorabies Virus with TK/gE Gene Deletion and Flt3L Co-Expression Enhances the Innate and Adaptive Immune Response via Activating Dendritic Cells

doi: 10.3390/v13040691

Figure Lengend Snippet: Characterization of the recombinant HNX-TK − /gE − -Flt3L. ( A ) Detection of the transcription of the Flt3L gene in HNX-TK − /gE − -Flt3L virus- and HNX-infected cells through RT-qPCR (**** < 0.001). ( B ) Detection of the Flt3L protein and viral gB protein expression through an immunofluorescence assay (IFA). Green: gB-positive cells; red: Flt3L-positive cells; blue: DAPI-stained PK-15 cell nucleus. DAPI, 4’,6-diamidino-2-phenylindole. ( C ) Western blot analysis of β-actin, gD, and Flt3L of the PK-15 cells infected with HNX-TK − /gE − -Flt3L, HNX, or DMEM as a control. ( D ) One step growth curves of HNX-TK − /gE − -Flt3L and HNX in PK-15 cells.

Article Snippet: The blots were blocked by skim milk (BD, Bioscience, Inc., Saint Louis, MO, USA), and this was followed by incubation with rabbit anti-Flt3L polyclonal antibody (Bioss, Beijing, China), mouse anti-gD monoclonal antibody (Keqian Ltd., Wuhan, China), and mouse anti-β-actin monoclonal antibody (Bioss).

Techniques: Recombinant, Infection, Quantitative RT-PCR, Expressing, Immunofluorescence, Staining, Western Blot

Journal: Journal of Nuclear Medicine

Article Title: 225 Ac/ 89 Zr-Labeled N4MU01 Radioimmunoconjugates as Theranostics Against Nectin-4–Positive Triple-Negative Breast Cancer

doi: 10.2967/jnumed.124.268387

Figure Lengend Snippet: Flow cytometry binding, internalization, and radioligand binding assay of anti–nectin-4 antibody (N4MU01) and its conjugates with (A) MDA-MB-231, MCF-7, MDA-MB-468, and 4T1. nectin-4 cells. Dose-dependent binding was observed in nectin-4–expressing cell lines, MDA-MB-468, MCF-7, and 4T1. nectin-4 . No binding was observed in MDA-MB-231 control cell line. (B) Internalization of N4MU01: MDA-MB-468, MCF-7, and MDA-MB-231 cells were treated with Incucyte FabFluor-labeled N4MU01 (4 μg/mL); High-definition phase and red fluorescence images (×10 magnification) were captured every 2 h for 48 h. All data are shown as mean ± SEM ( n = 3), and average internalization values for MCF-7 and MDA-MB-468 were compared with that of MDA-MB-231 using ANOVA. Estimation of K D values using radioligand binding assay for (C) [ 89 Zr]Zr-DFO-N4MU01 in nectin-4–positive MCF-7 and (D) [ 225 Ac]Ac-Macropa-N4MU01 in nectin-4–positive MDA-MB-468 cells. **** P < 0.0001. FITC-A = fluorescein isothiocyanate area; FL1-A = fluorescence intensity from channel 1 (green) area; FL2-A = fluorescence intensity from channel 2 (orange) area; PE-A = phycoerythrin area.

Article Snippet: The in vitro cytotoxicity (inhibitory concentration of 50% [IC 50 ]) of [ 225 Ac]Ac-Macropa-N4MU01, control immunoconjugates anti-CD20 [ 225 Ac]Ac-Macropa-rituximab, and unlabeled N4MU01 in nectin-4–positive MDA-MB-468, MCF-7, and nectin-4–negative MDA-MB-231 was determined using Incucyte Cytotox Red reagent in an Incucyte S3 live-cell imager (Essen BioScience) as previously reported (details in the supplemental materials) ( ).

Techniques: Flow Cytometry, Binding Assay, Radio Ligand Binding Assay, Expressing, Control, Labeling, Fluorescence

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: H2O2-induced different cell viability in M17, PC12 and MN9D cells as measured by MTT assay (A & B) and trypan blue dye exclusion assay (C). Each bar from Figs. represents data obtained from 5 separate experiments (N = 5 of independent cell culture preparations, which is the same in the following legends). *p < 0.05, **p < 0.01, ***p < 0.001, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: MTT Assay, Exclusion Assay, Cell Culture, Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on DDRs in M17 and MN9D cells as demonstrated by measurements of γH2AX (A) and p53 (B). Cells were exposed to 100–400 μM H2O2 for 3 h. Top panels: autoradiograph obtained by western blotting. Button panel: the quantitative analysis of band densities in western blotting. Each bar from Figs. represents data obtained from 5–7 separate experiments (N = 5–7). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Autoradiography, Western Blot, Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: H2O2 treatment induces single-strand DNA breaks as determined by the Comet assay in M17 and MN9D cells. Cells were exposed to different concentrations of H2O2 for 3 h. The cells were processed for comet assays run under alkaline conditions to identify DNA SSBs. Tail moment (B), tail length (C) and % DNA in the tail (D) were analyzed to evaluate DNA damage. Each bar from Figs. represents data obtained from 5 separate experiments (N = 5). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, compared to corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Single Cell Gel Electrophoresis, Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on ROS production in M17 and MN9D cells. Cells were exposed to different concentrations of for 3 h. ROS was measured by the DCFH2-DA assay. Each bar from Figs. represents data obtained from 6 separate experiments (N = 6). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to the corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on Ctr1 (A) and OGG1 (B) protein levels in M17 and MN9D cells. Each bar represents data obtained from 5 separate experiments (N = 5) in (A), and 4 separate experiments (N = 4) in (B). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to the corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Control

Journal: Neuroscience

Article Title: Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

doi: 10.1016/j.neuroscience.2019.09.036

Figure Lengend Snippet: Effects of H2O2 treatment on Cav1.2 (A) and Cav1.3 (B) protein levels in M17 and MN9D cells as determined by western blotting. Cells were exposed to different concentrations of H2O2 for 3 h. Each bar in Figs. represents data obtained from 6 to 7 separate experiments (N = 6–7). *p < 0.05, **p < 0.01, compared to the 0 group (control). †p < 0.05, ††p < 0.01, compared to the corresponding group in M17 cells.

Article Snippet: Cell cultures and drug exposure The growth medium for human neuroblastoma cell line SK-N-BE(2)-M17 (M17, ATCC, Cat#: CRL-2267) and the mouse hybrid dopaminergic cell line MN9D (transferred from Dr. Zigmond’s laboratory of University of Pittsburgh, MTA 0000434) was Dulbecco’s modified Eagle’s medium (DMEM), and for SH-SY5Y cells (ATCC, CRL-2266, Cat#: CRL-2266) was a 1:1 mix of DMEM and F12 media.

Techniques: Western Blot, Control