Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: Sequences of small-interfering RNAs

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques:

Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: Sequences of the primers used for RT-qPCR

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques: Sequencing

Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: Correlation analysis between LPS, inflammatory factors, and RAS pathway members. A Heat map showing the correlation between LPS, inflammatory cytokines and RAS members. Blue dots represent positive correlation and red dots represent negative correlation. Larger dots and darker colour correspond to greater correlation. Lower correlation is represented by smaller dots and lighter colour. B Ratio of ACE/ACE2 after treatment of BMEC with different concentrations of LPS. All data were represented as the mean ± SEM ( n = 3). ns, represents no significant difference ( p > 0.05); * represents p < 0.05; ** represents p < 0.01; and *** represents p < 0.001.

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques:

Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: Distribution of ACE2 in BMEC. Cells were stained with antibodies against ACE2 and nuclei were stained with 4’,6-diamidino-2-phenylindole (DAPI). Representative images were visualized by confocal laser microscopy. Red, ACE2; blue, DAPI. Scale bar: 10 μm.

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques: Staining, Microscopy

Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: DAD3 induces ACE2 expression in LPS-induced BMEC. BMEC were treated with LPS (0.5 μg/mL), DA (20 μg/mL), DAD3 (20 μg/mL), LPS+DA (0.5 μg/mL+20 μg/mL) and LPS+DAD3 (0.5 μg/mL+20 μg/mL) for 24 h. A Relative ACE2 mRNA levels were determined by RT-qPCR. B ACE2 protein expression levels were determined by Western blotting. β-actin was used as a control. All data were represented as the mean ± SEM ( n = 3). ns, represents no significant difference ( p > 0.05); * represents p < 0.05; ** represents p < 0.01; and *** represents p < 0.001.

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: Effects of ACE2 silencing and DAD3 treatments on the transcriptional or expression levels of RAS members. A-E BMEC were treated with either control siRNA (scrambled) or ACE2-siRNA for 24 h, followed by treatment with 0.5 μg/mL LPS for 24 h. After washing with PBS, cells were treated with 20 μg/mL DA or 20 μg/mL DAD3 for 24 h. The ACE2, AT1R and AT2R relative mRNA levels (transcriptional levels) of RAS members were determined by RT-qPCR. The Ang II and Ang-(1–7) protein expression levels of RAS members were determined by ELISA. A Relative ACE2 mRNA levels. B Expression levels of Ang II. C Expression levels of Ang-(1–7). D Relative AT1R mRNA levels. E Relative AT2R mRNA levels. All data were represented as the mean ± SEM ( n = 3). ns, represents no significant difference ( p > 0.05); * represents p < 0.05; ** represents p < 0.01; and *** represents p < 0.001.

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: Effects of ACE2 silencing and DAD3 treatments on the transcriptional levels of pro-inflammatory factors. A–D BMEC were treated with either control siRNA (scrambled) or ACE2-siRNA for 24 h, followed by treatment with 0.5 μg/mL LPS for 24 h. After washing with PBS, cells were treated with 20 μg/mL DA or 20 μg/mL DAD3 for 24 h. Relative mRNA levels of pro-inflammatory factors were determined by RT-qPCR. A Relative IL-1β mRNA levels. B Relative IL-6 mRNA levels. C Relative IL-8 mRNA levels. D Relative TNF-α mRNA levels. All data were represented as the mean ± SEM ( n = 3). ns, represents no significant difference ( p > 0.05); * represents p < 0.05; ** represents p < 0.01; and *** represents p < 0.001.

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques: Quantitative RT-PCR

Journal: Veterinary Research

Article Title: DAD3 targets ACE2 to inhibit the MAPK and NF-κB signalling pathways and protect against LPS-induced inflammation in bovine mammary epithelial cells

doi: 10.1186/s13567-022-01122-0

Figure Lengend Snippet: Effects of ACE2 silencing and DA and DAD3 treatment on the expression levels of components of the MAPK and NF-κB pathways. A, B BMEC were treated with either control siRNA (scrambled) or ACE2-siRNA for 24 h, followed by treatment with 0.5 μg/mL LPS for 24 h. After washing with PBS, cells were treated with 20 μg/mL DA or 20 μg/mL DAD3 for 24 h. The phosphorylation levels of p38, JNK1/2/3, ERK1/2 and IκB-α (p-p38, p-JNK, p-ERK1/2 and p-IκB-α) were determined by Western blotting. Activation of NF-κB pathway was indicated by p-IκB-α, an indicator for the activation of NF-κB. The band intensity of all detected proteins was normalized to β-actin and the expressions of p-p38, p-JNK, and p-ERK1/2 were normalized to p38, JNK, and ERK1/2, respectively. A DA treatments on the expression levels of components of the MAPK and NF-κB pathways; B DAD3 treatments on the expression levels of components of the MAPK and NF-κB pathways. All data were represented as the mean ± SEM ( n = 3). ns, represents no significant difference ( p > 0.05); * represents p < 0.05; ** represents p < 0.01; and *** represents p < 0.001.

Article Snippet: Anti-phospho-IκB-α (bs-2513R/Polyclonal/Rabbit), anti-ERK(bs-0022R/Polyclonal/Rabbit), anti-phospho-ERK(bs-1522R/Polyclonal/Rabbit), anti-JNK(bs-2592R/Polyclonal/Rabbit), anti-phospho-JNK (bs-1640R/Polyclonal/Rabbit), anti-ACE2 (bs-23444R/Polyclonal/Rabbit) and anti-β-actin (bs-0061R/Polyclonal/Rabbit) were from Bioss (Beijing, China).

Techniques: Expressing, Western Blot, Activation Assay

Journal: PLoS Pathogens

Article Title: SARS-CoV-2 variants with reduced infectivity and varied sensitivity to the BNT162b2 vaccine are developed during the course of infection

doi: 10.1371/journal.ppat.1010242

Figure Lengend Snippet: (A) Phylogenetic trees depicting N genes isolated from nose-throat swabs by SGS. IDs of individuals from whom swabs were isolated are indicated in the figure. Black rectangles depict the dominant N gene isolated from each swab sample. Red rectangles depict N genes with missense mutations, grey rectangles depict N genes with silent mutations and blue rectangles depict N genes with nonsense mutations. Mutations that are not documented in the GSAID are marked with an asterisk. For each N-gene variant, the nucleotide substitution is indicated (based on Wuhan-Hu-1 numbering). The segment with the number below each phylogenetic tree shows the length of branch that represents an amount genetic change. The amount of genetic change is the number of nucleotide substitutions divided by the length of the N-gene sequence. (B) Pie charts showing the proportion of all N-gene sequences isolated from each swab sample. The White pie slice depicts unmutated sequences. The Colored pie slices depict mutated N genes. The different mutations are indicated in the figure. The number in the middle of the pie chart depicts the total number of N-gene variants that were sequenced. (C) Relative p24 value as measured by adding 20 μl of supernatant containing the pseudovirus to Lenti-X GoStix Plus. The x-axis depicts the SARS-CoV-2 pseudoviruses that were produced, and the y-axis depicts the relative p24 protein (GoStix values) in the supernatant of each pseudovirus. (D) 293T-ACE2 infection with SARS-CoV-2 pseudovirses expressing mutated N proteins and unmutated (Wuhan-Hu-1) N protein (WT). The x-axis depicts the mutation in the N proteins of the pseudovirus that was used for infection. The y-axis depicts the luminescence levels that were measured 48 hours post infection. Experiments were done in triplicates and repeated three times. One representative experiment is shown. Mean values and standard errors are shown. Statistically significant differences in comparison to the WT SARS-CoV-2 pseudovirus are indicated (student’s t test, p < 0.005). Figure was generated using biorender.com .

Article Snippet: 293T-ACE2 (5x10 4 ) cells were seeded in a 96-well plate, washed, and were incubated with polyclonal anti-ACE2 antibody (Bioss, BS-23028R) or RBD-Ig [ ] for 1 hour at 4°C at 1:20 dilution, followed by incubation with PE-conjugated goat anti-human IgG (Jackson, Cat #109-116-088, 1:200) for 45 minutes.

Techniques: Isolation, Variant Assay, Sequencing, Produced, Infection, Expressing, Mutagenesis, Generated

Journal: PLoS Pathogens

Article Title: SARS-CoV-2 variants with reduced infectivity and varied sensitivity to the BNT162b2 vaccine are developed during the course of infection

doi: 10.1371/journal.ppat.1010242

Figure Lengend Snippet: (A) Schematic representation of the ten spike variants that were tested for infectivity. Amino acids substitution in each spike mutants is shown in the figure. NTD = N-terminal domain, RBD = receptor binding domain, FP = fusion peptide, HR1 = heptad repeat 1, HR2 = heptad repeat 2, TM = transmembrane domain, CT = cytoplasmic domain. (B) Relative p24 value as measured by adding 20 μl of supernatant containing the pseudovirus to Lenti-X GoStix Plus. The y-axis depicts the SARS-CoV-2 pseudoviruses that were produced, and the x-axis depicts the relative p24 protein (GoStix values) in the supernatant of each pseudovirus. WT = pseudovirus that expresses an unmutated (Wuhan-Hu-1) spike. (C) FACS staining of 293T-ACE2 cells. The gray histogram shows the staining of the 293T-ACE2 cells with secondary antibody only. The empty black histograms depict the staining anti-ACE2 antibody or RBD-Ig. Shown is one representative experiment out of three preformed. (D) 293T-ACE2 infection with SARS-CoV-2 pseudovirses expressing mutated spikes, unmutated (Wuhan-Hu-1) spike protein and bald pseudovirus. The x-axis depicts the mutation in the spike of the pseudovirus that was used for infection. The y-axis depicts the luminescence levels that were measured 48 hours post infection. Experiments were done in triplicates and repeated three times. One representative experiment is shown. Mean values and standard errors are shown. WT = pseudovirus that expresses an unmutated (Wuhan-Hu-1) spike. Statistically significant differences in comparison to the WT SARS-CoV-2 pseudovirus are indicated (student’s t test, *p < 0.05, **p < 0.005). Figure was generated using biorender.com .

Article Snippet: 293T-ACE2 (5x10 4 ) cells were seeded in a 96-well plate, washed, and were incubated with polyclonal anti-ACE2 antibody (Bioss, BS-23028R) or RBD-Ig [ ] for 1 hour at 4°C at 1:20 dilution, followed by incubation with PE-conjugated goat anti-human IgG (Jackson, Cat #109-116-088, 1:200) for 45 minutes.

Techniques: Infection, Binding Assay, Produced, Staining, Expressing, Mutagenesis, Generated

Journal: PLoS Pathogens

Article Title: SARS-CoV-2 variants with reduced infectivity and varied sensitivity to the BNT162b2 vaccine are developed during the course of infection

doi: 10.1371/journal.ppat.1010242

Figure Lengend Snippet: (A, C) OD values of ELISA against spike S1 subunit of plasma from vaccinated individuals (A) and of convalescent plasma (C). Experiments were done in triplicates and repeated three times. One representative experiment is shown. Mean values and standard errors are shown. OD values of control plasma (unvaccinated healthy individual) are shown in the red dashed graph. (B) The luminescence values derived from 293T-ACE2 cells 48 hours after infection with nanoluc-expressing SARS-CoV-2 pseudovirus and following incubation with increasing concentrations of plasma from vaccinated individuals. The plasma dilution is shown in the x-axis. Luminescence values after incubation with control (unvaccinated healthy individual) is shown in the red-dashed graph. Experiments were done in triplicates and repeated two times. Mean values and standard errors are shown; representative of two independent experiments is shown. (D) The luminescence values derived from 293T-ACE2 cells 48 hours post infection with nanoluc-expressing SARS-CoV-2 pseudovirus and following incubation with convalescent plasma at 10 −3 dilution. The study ID of the plasma samples is shown in the x-axis. Experiments were done in triplicates and repeated two times. Mean values and standard errors are shown; representative of two independent experiments is shown.

Article Snippet: 293T-ACE2 (5x10 4 ) cells were seeded in a 96-well plate, washed, and were incubated with polyclonal anti-ACE2 antibody (Bioss, BS-23028R) or RBD-Ig [ ] for 1 hour at 4°C at 1:20 dilution, followed by incubation with PE-conjugated goat anti-human IgG (Jackson, Cat #109-116-088, 1:200) for 45 minutes.

Techniques: Enzyme-linked Immunosorbent Assay, Derivative Assay, Infection, Expressing, Incubation

Journal: PLoS ONE

Article Title: Endoplasmic Reticulum Stress-Sensing Mechanism Is Activated in Entamoeba histolytica upon Treatment with Nitric Oxide

doi: 10.1371/journal.pone.0031777

Figure Lengend Snippet: Genes upregulated encoding metabolism enzymes.

Article Snippet: The primary antibody used were a rabbit anti-PFOR polyclonal antibody (1∶400 dilution, a kind gift from Dr Ester Orozco, CINVESTAV, Mexico), a mouse anti-actin monoclonal antibody (1∶20,000 dilution; #69100, MP Biomedicals).

Techniques:

Journal: PLoS ONE

Article Title: Endoplasmic Reticulum Stress-Sensing Mechanism Is Activated in Entamoeba histolytica upon Treatment with Nitric Oxide

doi: 10.1371/journal.pone.0031777

Figure Lengend Snippet: PFOR of 120 kDa was revealed upon SDS-PAGE and immunobloting of crude extracts from SNP-treated and control trophozoites (4, 2 and 1×10 4 cells). The loaded amount of proteins for each condition is evidenced by actin (43 kDa) immunodetection on the same western blot. Quantification of signal emission did not reveal differences between the tested conditions in 3 independent experiments.

Article Snippet: The primary antibody used were a rabbit anti-PFOR polyclonal antibody (1∶400 dilution, a kind gift from Dr Ester Orozco, CINVESTAV, Mexico), a mouse anti-actin monoclonal antibody (1∶20,000 dilution; #69100, MP Biomedicals).

Techniques: SDS Page, Western Blot, Control, Immunodetection

Journal: PLoS ONE

Article Title: Endoplasmic Reticulum Stress-Sensing Mechanism Is Activated in Entamoeba histolytica upon Treatment with Nitric Oxide

doi: 10.1371/journal.pone.0031777

Figure Lengend Snippet: PFOR activities upon no treatment.

Article Snippet: The primary antibody used were a rabbit anti-PFOR polyclonal antibody (1∶400 dilution, a kind gift from Dr Ester Orozco, CINVESTAV, Mexico), a mouse anti-actin monoclonal antibody (1∶20,000 dilution; #69100, MP Biomedicals).

Techniques: Activity Assay