incucyte zoom  (Sartorius AG)

Journal: PLOS One

Article Title: Omicron SARS-CoV-2 shows NRP1-linked susceptibility in adipose-derived MSCs and alters primitive cord blood CD34⁺ HSPCs

doi: 10.1371/journal.pone.0360439

Figure Lengend Snippet: (A) Detection of SARS-CoV-2 infected cells using high-content imaging under control conditions. Fluorescent signals: green (anti-SARS-CoV NP mAb), blue (Hoechst). (B) Representative flow cytometry analysis plots showing NP+ cells. (C) Measurement of fluorescent intensity in NP+ cells. (D) Percentage of infected MSCs. (E) The supernatants of infected MSCs were collected and titrated on Vero-E6/TMPRSS2 cells to measure infectious viral particle production using the focus-forming assay. (F) After RNA extraction from infected MSCs, the expression of IDO1 was examined using real-time PCR. The graph displays the expression of IDO1 normalized to GAPDH , with the mock group serving as the control. Data are represented as means ± SEM from three independent biological replicates. For , the Delta condition was available from only two independent donors and was therefore presented descriptively and excluded from the statistical analysis. Statistical comparisons among the remaining groups were performed using one-way ANOVA followed by Tukey’s multiple-comparisons test. One-way ANOVA showed a significant overall difference among groups, F(2, 6) = 6.794, P = 0.0287. * P < 0.05, ** P < 0.01, *** P = 0.0001, **** P < 0.0001.

Article Snippet: The Operetta high-content imaging system (PerkinElmer) was used to detect and analyze the fluorescent signals.

Techniques: Infection, Imaging, Control, Flow Cytometry, Analysis, Focus Forming Assay, RNA Extraction, Expressing, Real-time Polymerase Chain Reaction

Journal: PLOS One

Article Title: Omicron SARS-CoV-2 shows NRP1-linked susceptibility in adipose-derived MSCs and alters primitive cord blood CD34⁺ HSPCs

doi: 10.1371/journal.pone.0360439

Figure Lengend Snippet: (A) Representative western blot depicting the expression of NRP1 in AD-MSCs (No siRNA), cells transfected with non-targeting siRNA (siControl), or siNRP1 for 24 hours. β-actin was used as a loading control. (B) Graph displaying the quantification of NRP1 normalized to β-actin (n = 3). This experiment represents three independent biological replications. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. (C) Transfected cells were visualized using high-content imaging. Fluorescent signals: green for anti-NRP1 mAb, blue for Hoechst. (D) Percentage of NRP1-expressing cells normalized to cells transfected with siControl. Data are presented as mean ± SEM from three independent experiments. Statistical analysis was conducted using a paired t-test. Next, siRNA-transfected cells were infected with different variants of SARS-CoV-2. (E) SARS-CoV-2-infected cells were detected using high-content imaging. Fluorescent signals: green for anti-SARS-CoV NP mAb, blue for Hoechst. (F) The fluorescent intensity of NP+ cells was measured. All data represent three independent experiments. Statistical analysis among each group of variant was performed using an unpaired t-test. * P < 0.05, ** P < 0.01, *** P = 0.0001, **** P < 0.0001.

Article Snippet: The Operetta high-content imaging system (PerkinElmer) was used to detect and analyze the fluorescent signals.

Techniques: Western Blot, Expressing, Transfection, Control, Analysis, Imaging, Infection, Variant Assay