Journal: Thoracic Cancer

Article Title: Circ_0001667 accelerates breast cancer proliferation and angiogenesis through regulating CXCL10 expression by sponging miR ‐6838‐5p

doi: 10.1111/1759-7714.14820

Figure Lengend Snippet: Knockdown of circ_0001667 inhibited proliferation and angiogenesis in breast cancer cells. MDA‐MB‐231 and BT549 cells were transfected with si‐NC or si‐circ_0001667. (a) qRT‐PCR assay for silencing efficiency of si‐circ_0001667. (b‐d) CCK8 and EdU assays to detect proliferation of breast cancer cells. Scale bar: 50 μm. (e and f) Flow cytometry was used to determine the cycle distribution of breast cancer cells. (g) Colony formation assay was used to assess the colony formation ability. (h) Western blot to measure the protein levels of PCNA and Ki67. (i) Tube formation assay was used to analyze angiogenesis of breast cancer cells. (j) Western blot showing the expression levels of VEGFA and FGF2 in breast cancer cells. * p < 0.05.

Article Snippet: The separated proteins were electrotransferred to PVDF membrane (Beyotime), closed with 10% skimmed milk, and incubated with primary antibodies (Abcam) against PCNA (ab18197, 1:1000), Ki67 (ab254123, 1:5000), VEGFA (ab46154, 1:2000), FGF2 (ab208687, 1:1000), CXCL10 (ab9807, 1:2000), or β‐actin (ab6276, 1:5000).

Techniques: Knockdown, Transfection, Quantitative RT-PCR, Flow Cytometry, Colony Assay, Western Blot, Tube Formation Assay, Expressing

Journal: Thoracic Cancer

Article Title: Circ_0001667 accelerates breast cancer proliferation and angiogenesis through regulating CXCL10 expression by sponging miR ‐6838‐5p

doi: 10.1111/1759-7714.14820

Figure Lengend Snippet: Downregulation of miR‐6838‐5p attenuated the effect of circ_0001667 knockdown on cell proliferation and angiogenesis. (a) qRT‐PCR analysis of miR‐6838‐5p levels in cells transfect with anti‐miR‐6838‐5p or anti‐miR‐NC. (b‐j) MDA‐MB‐231 and BT549 cells were transfected with si‐NC, si‐circ_0001667, si‐circ_0001667 + anti‐miR‐NC or si‐circ_0001667 + anti‐miR‐6838‐5p. (b‐d) CCK‐8 and EdU assays detected cell proliferation. (e and f) Flow cytometry determined cell cycle distribution. (g) The colony formation assay assessed the colony formation ability. (h) Western blot measured the protein levels of PCNA and Ki67. (i) Tube formation test analyzed angiogenesis. (j) Western blot showed the expression levels of VEGFA and FGF2. * p < 0.05.

Article Snippet: The separated proteins were electrotransferred to PVDF membrane (Beyotime), closed with 10% skimmed milk, and incubated with primary antibodies (Abcam) against PCNA (ab18197, 1:1000), Ki67 (ab254123, 1:5000), VEGFA (ab46154, 1:2000), FGF2 (ab208687, 1:1000), CXCL10 (ab9807, 1:2000), or β‐actin (ab6276, 1:5000).

Techniques: Knockdown, Quantitative RT-PCR, Transfection, CCK-8 Assay, Flow Cytometry, Colony Assay, Western Blot, Expressing

Journal: Thoracic Cancer

Article Title: Circ_0001667 accelerates breast cancer proliferation and angiogenesis through regulating CXCL10 expression by sponging miR ‐6838‐5p

doi: 10.1111/1759-7714.14820

Figure Lengend Snippet: Overexpression of CXCL10 reversed the effects of miR‐6838‐5p overexpression on cell proliferation and angiogenesis. (a and b) Validation of CXCL10 overexpression by qRT‐PCR and western blot. (c–k) MDA‐MB‐231 and BT549 cells were transfected with miR‐NC, miR‐6838‐5p, miR‐6838‐5p + pcDNA or miR‐6838‐5p + CXCL10. (c–h) CCK8 assay, EdU assay, flow cytometry and colony formation assay detected cell proliferation. (i) Western blot analysis for PCNA and Ki67 levels. (j) Tube formation test detected angiogenesis. (k) Western blotting quantified the levels of VEGFA and FGF2. * p < 0.05.

Article Snippet: The separated proteins were electrotransferred to PVDF membrane (Beyotime), closed with 10% skimmed milk, and incubated with primary antibodies (Abcam) against PCNA (ab18197, 1:1000), Ki67 (ab254123, 1:5000), VEGFA (ab46154, 1:2000), FGF2 (ab208687, 1:1000), CXCL10 (ab9807, 1:2000), or β‐actin (ab6276, 1:5000).

Techniques: Over Expression, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Transfection, CCK-8 Assay, EdU Assay, Flow Cytometry, Colony Assay

Journal: iScience

Article Title: Influenza A virus infection activates STAT3 to enhance SREBP2 expression, cholesterol biosynthesis, and virus replication

doi: 10.1016/j.isci.2024.110424

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-NS1 , Santa Cruz , Cat# sc-130568; RRID: AB_2011757.

Techniques: Virus, Recombinant, Staining, RNA Extraction, cDNA Synthesis, Transfection, Cloning, Cell Viability Assay, Software, CRISPR

Journal: Science translational medicine

Article Title: Nonhuman primates exposed to Zika virus in utero are not protected against reinfection at one year postpartum

doi: 10.1126/scitranslmed.aaz4997

Figure Lengend Snippet: Shown is the study timeline for ZIKV infection of rhesus macaque dams, delivery of offspring, blood and cerebrospinal fluid sampling, MRI, reinfection of offspring with ZIKV at 13–15 months of age, and necropsy. M, mock-infected; E, early gestational infection; L, late gestational infection; D, dam; I, infant offspring. The first dam and infant pair from each cohort were euthanized at the time of delivery and their tissues were analyzed. The remainder of the infants were observed for the entirety of the study.

Article Snippet: The purified recombinant ZIKV proteins (ZIKV-E from Sino Biologicals and ZIKV-NS1 from Meridian Life Sciences) were coupled to a GLC sensor chip via amine coupling with either 100 or 500 resonance units (RU) in the test flow channels.

Techniques: Infection, Sampling

Journal: Science translational medicine

Article Title: Nonhuman primates exposed to Zika virus in utero are not protected against reinfection at one year postpartum

doi: 10.1126/scitranslmed.aaz4997

Figure Lengend Snippet: Serum and amniotic fluid samples were obtained prenatally and postnatally at different time points after infection of macaque dams with ZIKV. Samples were assayed for ZIKV RNA (Log10 GE/ml) by qPCR; negative ZIKV RNA readings are not shown. Total ZIKV-specific neutralizing antibodies in serum are represented by the dilution that achieved a 50% reduction in viral plaques (PRNT50 test). All PRNT50 positive and negative readings are shown. The vertical dotted line indicates time of birth for offspring. All ZIKV-infected dam-infant pairs are shown. All ZIKV RNA and PRNT50 measurements from the mock-infected dam-infant pairs were below the limit of detection.

Article Snippet: The purified recombinant ZIKV proteins (ZIKV-E from Sino Biologicals and ZIKV-NS1 from Meridian Life Sciences) were coupled to a GLC sensor chip via amine coupling with either 100 or 500 resonance units (RU) in the test flow channels.

Techniques: Infection

Journal: Science translational medicine

Article Title: Nonhuman primates exposed to Zika virus in utero are not protected against reinfection at one year postpartum

doi: 10.1126/scitranslmed.aaz4997

Figure Lengend Snippet: Shown are the kinetics of IgM, IgA, and IgG antibodies against ZIKV envelope (E) protein and NS1 protein in blood samples from ZIKV-infected dams (A) and their infant offspring (B). Antibody binding to ZIKV protein-coated sensors was detected by surface plasmon resonance and is expressed as maximum relative units (Max RU). Max RU was calculated by multiplying the observed RU signal by dilution factors. The dashed line indicates the lower limit of detection for the assay. E, early gestational infection; L, late gestational infection; D, dam; I, infant offspring.

Article Snippet: The purified recombinant ZIKV proteins (ZIKV-E from Sino Biologicals and ZIKV-NS1 from Meridian Life Sciences) were coupled to a GLC sensor chip via amine coupling with either 100 or 500 resonance units (RU) in the test flow channels.

Techniques: Infection, Binding Assay, SPR Assay

Journal: Science translational medicine

Article Title: Nonhuman primates exposed to Zika virus in utero are not protected against reinfection at one year postpartum

doi: 10.1126/scitranslmed.aaz4997

Figure Lengend Snippet: Two cohorts of animals were used in this study: the first comprised offspring born to dams that were not infected with ZIKV during pregnancy (15-month-old M2I and two additional two-year-old animals, C1 and C2), and the second cohort comprised the four 13–15-month-old offspring (E2I, E3I, L2I, L3I) born to dams that were infected with ZIKV while pregnant. All seven macaques were inoculated with the same strain and dose of ZIKV used for the primary infection (green triangle). Blood samples were taken at 1, 2, 3, 4, 5, 7, 10 and 12 days after inoculation (blue triangles). Cerebrospinal fluid was collected on days −14 (2 weeks before reinfection), 3 and 7 days after reinfection and at necropsy (blue triangles). Necropsies were performed 14–16 days after ZIKV reinfection. M, a macaque infant born to a dam mock-infected while pregnant; E, macaque infants born to dams infected with ZIKV during early gestation; L, macaque infants born to dams infected with ZIKV during late gestation; I, infant offspring.

Article Snippet: The purified recombinant ZIKV proteins (ZIKV-E from Sino Biologicals and ZIKV-NS1 from Meridian Life Sciences) were coupled to a GLC sensor chip via amine coupling with either 100 or 500 resonance units (RU) in the test flow channels.

Techniques: Infection

Journal: Science translational medicine

Article Title: Nonhuman primates exposed to Zika virus in utero are not protected against reinfection at one year postpartum

doi: 10.1126/scitranslmed.aaz4997

Figure Lengend Snippet: (A) ZIKV RNA (Log10 GE/ml) was quantified by qPCR in serum and cerebrospinal fluid (CSF) of M2I, C1 and C2 macaques (cohort 1, no prior ZIKV exposure), and E2I, E3I, L2I, L3I (cohort 2, 13–15-month-old offspring born to dams infected with ZIKV while pregnant). Results for all serum samples are shown. Only positive CSF measurements are indicated. The dotted line marks the limit of detection of the assay. (B) Shown are the amounts of total ZIKV-specific neutralizing antibodies in blood samples from the seven macaques represented by the dilution that achieved a 50% reduction in viral plaques (PRNT50 test). All PRNT50 data are shown. (C) Quantification by droplet digital PCR of ZIKV RNA in lymphoid, pulmonary, digestive, genitourinary and nervous system tissues from the two cohorts is presented. (D) Shown are ZIKV-specific T cell memory responses in blood samples from the seven macaques as well as a single dam infected during early pregnancy (E2D) measured by flow cytometry. Infant samples (E2I, E3I, L2I, L3I) were collected 30 days before the reinfection study; the dam sample was collected 23 days after inoculation with ZIKV during pregnancy. Data are presented as the percentage of cytokine-positive T cells after stimulation of PBMCs with ZIKV envelope and NS1 peptides (with subtraction of baseline, which is the percentage of the same cells producing cytokines without stimulation). Statistically significant differences between the peptide stimulated (N=3 technical replicates for each cell/cytokine combination) and unstimulated groups (N=3 technical replicates) were determined by Student’s t test (*p<0.05, **p<0.01, ***p<0.001). Data shown are mean +/− SEM. (E) Shown is a representative thoracic spinal cord section from infant E3I stained with hematoxylin and eosin showing mineralized lesions after ZIKV reinfection. Wallerian degeneration associated with digestion chambers in the cytoplasm of Schwann cells is indicated by black arrows. (F) Shown is a representative normal thoracic spinal cord section from infant M2I stained with hematoxylin and eosin. Scale bar is 20 μm.

Article Snippet: The purified recombinant ZIKV proteins (ZIKV-E from Sino Biologicals and ZIKV-NS1 from Meridian Life Sciences) were coupled to a GLC sensor chip via amine coupling with either 100 or 500 resonance units (RU) in the test flow channels.

Techniques: Infection, Digital PCR, Flow Cytometry, Staining

Journal: Biosensors & bioelectronics

Article Title: The capacitive sensing of NS1 Flavivirus biomarker.

doi: 10.1016/j.bios.2016.08.097

Figure Lengend Snippet: Fig. 2. ECS-ES approach for NS1 protein detection in PBS using the anti-NS1 functionalized electrode. (a) Example of Nyquist capacitive plots (C′′ is the imaginary and C′ is the real part of capacitance) recorded as a function of different concentrations of NS1 in PBS. Cr was measured by the projection of the semicircle diameter in the axis C′, corresponding to the frequency value where C′′ is minimized (at low frequencies) as shown in the inset. (b) Calibration curve using logarithm of different concentrations of NS1 in PBS (R2∼0.996) wherein the RSD showed for each concentration (errors bars) represents three repetitions of the same functionalized electrode. Note that this analytical curve was obtained by plotting RR C %∆[1/ ] r =100×( C 1/ r NS ( 1)−C 1/ r blank ( ))/ C 1/ ) r blank ( ) , where C 1/ r blank , is the inverse of the capacitance of the biosensor after exposing its surface to PBS solution without a target. Subsequently the surface was evaluated against different concentrations of NS1 (plotted in logarithm).

Article Snippet: All reagents described in this work were purchased from SigmaAldrich, except the recombinant dengue virus NS1 glycoprotein (ab64456), monoclonal antibody anti-NS1, IgG1 isotype (ab138696) that were purchased from Abcam.

Techniques: Concentration Assay

Journal: Biosensors & bioelectronics

Article Title: The capacitive sensing of NS1 Flavivirus biomarker.

doi: 10.1016/j.bios.2016.08.097

Figure Lengend Snippet: Fig. 3. EIS-ES assay for NS1 protein in PBS. (a) Nyquist impedimetric plots recorded at different concentrations of NS1; the applied potential was ∼0.22 V. Rct was calculated by fitting the data to the Randle-like equivalent circuit model as illustrated in the inset in (a). The red lines are the adjusted curves to the equivalent circuit model. (b) Analytical calibration curve (R2∼0.990) using RR% against logarithm of NS1 at different concentrations with RSD (%) represented by error bars. Results extracted and adapted with permission from Cecchetto et al. (2015).

Article Snippet: All reagents described in this work were purchased from SigmaAldrich, except the recombinant dengue virus NS1 glycoprotein (ab64456), monoclonal antibody anti-NS1, IgG1 isotype (ab138696) that were purchased from Abcam.

Techniques:

Journal: Biosensors & bioelectronics

Article Title: The capacitive sensing of NS1 Flavivirus biomarker.

doi: 10.1016/j.bios.2016.08.097

Figure Lengend Snippet: Fig. 4. Response of anti-NS1 interface in the presence of a nonspecific protein (Fetuin). a) Inverse of the capacitive response was typically less than 5% of C ∆(1/ ) r , in contrast to a variation of 43% of C ∆(1/ ) r for the specific response. b) Impedimetric response was typically less than 6% of Rct blockage, in contrast to a variation of 41% of Rct blockage for the specific response. Figure (b) reproduced with permission from Cecchetto et al. (2015).

Article Snippet: All reagents described in this work were purchased from SigmaAldrich, except the recombinant dengue virus NS1 glycoprotein (ab64456), monoclonal antibody anti-NS1, IgG1 isotype (ab138696) that were purchased from Abcam.

Techniques:

Journal: Biosensors & bioelectronics

Article Title: The capacitive sensing of NS1 Flavivirus biomarker.

doi: 10.1016/j.bios.2016.08.097

Figure Lengend Snippet: Fig. 5. Detection of NS1 protein in neat serum by anti-NS1 functionalized electrode using ECS and EIS sensorial approaches. (a) Calibration curve using C 1/ r (ECS-ES) as signal transducer (R2 of 0.98) as a function of logarithm of NS1 at different concentrations, where RR C %∆[1/ ] r =100×( C 1/ r NS ( 1)−C 1/ r blank ( ))/ C 1/ ) r blank ( ) . The error bars indicate the standard deviation of triplicate measurements. (b) Calibration curve using Rct (EIS-ES) as signal transducer (R2 of 0.98 as a function of logarithm of NS1 at different concentrations Figure “b” adapted with permission from Cecchetto et al. (2015).

Article Snippet: All reagents described in this work were purchased from SigmaAldrich, except the recombinant dengue virus NS1 glycoprotein (ab64456), monoclonal antibody anti-NS1, IgG1 isotype (ab138696) that were purchased from Abcam.

Techniques: Standard Deviation

Journal: Emerging microbes & infections

Article Title: Extracellular acidosis enhances Zika virus infection both in human cells and ex-vivo tissue cultures from female reproductive tract.

doi: 10.1080/22221751.2021.1932606

Figure Lengend Snippet: Figure 2. Acidosis enhances ZIKV infection in cells derived from the female reproductive tract and other target cells. Cells were incubated with ZIKV (A1C1-V2) at different pH values for 1 h at 37°C. Then, cells were washed and infection was assessed 72 h later. Infection of genital tract-derived cells VK2/E6E7 (A), Ect1/E6E7 (B), and End1/E6E7 (C) was quantified by measuring the extra- cellular concentration of NS-1 by Luminex. Infection of human endometrial fibroblasts (T-HESC) was quantified by qPCR (D). Infec- tion of monocyte-derived macrophages (MDM) was quantified by measuring the extracellular concentration of NS-1 by Luminex (E). Primary culture-derived human astrocytes (NHA) infection was measured by qPCR (F). Human ectocervical (G) and endocervical (H) tissue cultures were dissected into 2 mm [3] pieces and infected with ZIKV (VR-1844 moi 0.1) either at pH 7.3 or 6.2 for 1 h at 37°C. The viral production was evaluated at days 3, 6, 10, 13, 17, and 21 by measuring ZIKV NS1 in the culture supernatant. Results in A, B, C, D, and F represent data of two independent experiments. Data in E comprises the results of four different donors. Panels H and G were constructed with data of three independent experiments per explant type, and data were analysed using paired two-way ANOVA. Results are presented as mean ± SEM.

Article Snippet: The standard curve was obtained by serial dilution of recombinant NS1 protein (Fitzgerald, cat no. 30-1931).

Techniques: Infection, Derivative Assay, Incubation, Concentration Assay, Luminex, Construct

Journal: Diagnostics

Article Title: Development of a Flow-Free Automated Colorimetric Detection Assay Integrated with Smartphone for Zika NS1

doi: 10.3390/diagnostics10010042

Figure Lengend Snippet: ( a ) Sandwich ELISA assay results using the recombinant Zika nonstructural protein 1 (NS1) antigen spiked on PBS buffer on 96-well plate as the Zika NS1 monoclonal antibody is used as a capture agent. Anti-Zika NS1 monoclonal antibody-HRP with a dilution factor of 1:1000 utilized to react with TMB calorigenic substrate to develop color. ( b ) M-ELISA assay on a 96-well plate showing spiking recombinant Zika NS1 on whole plasma. A 1:500 dilution factor of HRP-labeled anti-Zika NS1 was used and reacted with TMB to generate color. In both cases, color development was stopped by using H 2 SO 4 and absorbance was measured at 450 nm using a SpectraMax Gemini™ XPS/EM Microplate Reader (Molecular Devices, USA). Error bars are ±SD.

Article Snippet: It was rewashed three times carefully, followed by 90 min incubation of 100 μL of recombinant Zika NS1 antigen (BF-NS1-6309, Biofront Technologies, Tallahassee, Florida, USA) with different concentrations spiked in PBS at room temperature.

Techniques: Sandwich ELISA, Recombinant, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Labeling