Journal: Virulence

Article Title: Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19

doi: 10.1080/21505594.2022.2073025

Figure Lengend Snippet: Prevalence of IgG specific for betacoronavirus RBDs in study cohort. Plasma was assayed by ELISA using recombinant, multimeric RBD as antigen. (a) SARS-COV-2 spike 319-591 with p values from ANOVA with Tukey HSD for all pairwise comparisons shown beneath. (b) SARS-CoV spike 384-655 with p values shown as above, (c) MERS spike 306-577. Each point indicates an individual participant. Blue bars show median values. N = 7–30. (d) Comparison of absorbance values for MERS and SARS-CoV RBD binding IgG for all participants. Each point indicates an individual participant, n = 124. The orange circle indicates participants with detectable amounts of MERS RBD-binding IgG, showing correlation with SARS-CoV-binding IgG in this subset. IgG specific to recombinant, multimeric RBD from seasonal betacoronaviruses, (e) OC43 spike 315-675, (f) HKU1 spike 307-675 assayed by ELISA as above.

Article Snippet: ELISAs were prepared using the following recombinant antigens (expressed in HEK 293 cells with 8 × His tags) coated onto 96-well plates: SARS-CoV-2 Spike RBD (multimeric, 319–591, Cell Signaling Technology #17862), SARS-CoV RBD (multimeric, 306–577), MERS-CoV RBD (multimeric, 364–655), OC43 Spike RBD (multimeric, 315–675), HKU1 Spike RBD (multimeric, 307–675).

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Binding Assay

Journal: Virulence

Article Title: Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19

doi: 10.1080/21505594.2022.2073025

Figure Lengend Snippet: Prevalence of IgGs specific for SARS-CoV-2 peptides. (b) Individual peptides with significant variation between groups ( p < 0.05, ANOVA, n = 7–30). Bars denote groups with significant differences by pairwise comparison (Tukey HSD, p < 0.05).

Article Snippet: ELISAs were prepared using the following recombinant antigens (expressed in HEK 293 cells with 8 × His tags) coated onto 96-well plates: SARS-CoV-2 Spike RBD (multimeric, 319–591, Cell Signaling Technology #17862), SARS-CoV RBD (multimeric, 306–577), MERS-CoV RBD (multimeric, 364–655), OC43 Spike RBD (multimeric, 315–675), HKU1 Spike RBD (multimeric, 307–675).

Techniques:

Journal: Virulence

Article Title: Repertoires of SARS-CoV-2 epitopes targeted by antibodies vary according to severity of COVID-19

doi: 10.1080/21505594.2022.2073025

Figure Lengend Snippet: Comparison between peptide seropositivity frequencies in the primary dataset with an independent cohort. Peptides that showed >20 percentage point difference in frequency of seropositivity between participants in the Chelsea survey who had been hospitalized and those who reported exposure to SARS-CoV-2 are shown in green. The differences in frequency of seropositivity for IgG specific to those peptides between acutely hospitalized patients and people with no history of infection are shown in blue.

Article Snippet: ELISAs were prepared using the following recombinant antigens (expressed in HEK 293 cells with 8 × His tags) coated onto 96-well plates: SARS-CoV-2 Spike RBD (multimeric, 319–591, Cell Signaling Technology #17862), SARS-CoV RBD (multimeric, 306–577), MERS-CoV RBD (multimeric, 364–655), OC43 Spike RBD (multimeric, 315–675), HKU1 Spike RBD (multimeric, 307–675).

Techniques: Infection

Journal: Molecular Therapy. Nucleic Acids

Article Title: circRNA CDR1as Promotes Pulmonary Artery Smooth Muscle Cell Calcification by Upregulating CAMK2D and CNN3 via Sponging miR-7-5p

doi: 10.1016/j.omtn.2020.09.018

Figure Lengend Snippet: Verification of CAMK2D (CA) and CNN3 (CN) but Not CACNB4 and CACNG7 as Cognate Targets of miR-7-5p (A) TargetScan-predicted binding sites of targets with miR-7-5p. (B) Effects of miR-7-5p on the expression levels of CACNB4, CACNG7, CAMK2D, and CNN3. ∗p < 0.05, ∗∗p < 0.01 versus Nor + NC; #p < 0.05, ##p < 0.01 versus Hyp + NC. (C) Diagrammatic sketch of the binding sites for the WT and MUT binding sites of CAMK2D and CNN3 associated with miR-7-5p. (D) Luciferase reporter assay for the luciferase activity of HPASMCs cotransfected with scrambled miRNA (NC) and VE/CAMK2D and CNN3 3′ UTR-WT/CAMK2D and CNN3 3′ UTR-MUT/complementary sequences of miR-7-5p (miR-7-5p-PC) or miR-7-5p mimics with VE/CAMK2D and CNN3 3′ UTR-WT/CAMK2D and CNN3 3′ UTR-MUT/complementary sequences of miR-7-5p (miR-7-5p-PC). ∗∗p < 0.01 compared with NC + WT; ##p < 0.01 compared with NC + miR-7-5p-PC. (E and F) HPASMCs were labeled with antibodies against CNN3 (E) and CAMK2D (F), and nuclei were stained with DAPI and merged to represent an overlay figure. Scale bars, 25 μm. All tests were performed at least three times, and the values are presented as the mean ± SEM.

Article Snippet: The antibodies and reagents used were as follows: Runx2 (ab23981; Abcam, MA, USA); MSX2 (M-70; sc-15396; Santa Cruz Biotechnology, TX, USA); BMP2 (ab14933; Abcam, MA, USA), SOX9 (ab26414; Abcam, MA, USA); SM22α (ab14106; Abcam, MA, USA), CACNB4 (17770-1-AP; Proteintech, IL, USA); CACNG7 (17862-1-AP; Proteintech, IL, USA); CAMK2D (20667-1-AP; Proteintech, IL, USA, and ab52476; Abcam, MA, USA); CNN3 (11509-1-AP; Proteintech, IL, USA); ARS (GMS80046; GENMED, Shanghai, PR China); and the ALP Assay Kit (P0321; Beyotime, Shanghai, PR China).

Techniques: Binding Assay, Expressing, Luciferase, Reporter Assay, Activity Assay, Labeling, Staining