snp ace2 c 2551626 1  (Thermo Fisher)

Journal: MedComm

Article Title: Blocking Extracellular Cyclophilin A‐Mediated Adhesion Protects Against Bacterial Infection

doi: 10.1002/mco2.71063

Figure Lengend Snippet: Binding of eCypA to TβR2 facilitates the formation of the TGFβ1‐TGF‐β receptor 2 complex (A) qPCR analysis of ITGA5 and FN mRNA levels in A549 cells transfected with siRNAs targeting TβR2, CD147, ACE2, or MUC18 for 48 h, and then treated with 10 nM eCypA or PBS for 12 h. (B) Immunoblot analysis of the indicated proteins in immunoprecipitated samples and whole‐cell lysates (WCL) of A549 cells treated with Fc, CypA‐Fc (200 ng/mL), R55‐Fc (200 ng/mL), CypA‐Fc + anti‐CypA mAb (1 µg/mL), or R55‐Fc + anti‐CypA mAb for 2 h. (C) Interactions of CypA with TGF‐β1, TβR1, and TβR2 were detected using ELISA. BSA was used as a negative control. (D) The blocking effect of the anti‐CypA mAb on the binding of CypA to TβR2 was detected using ELISA. (E–H) Interaction of CypA with anti‐CypA mAb (E) and TβR2 (F) was detected by SPR. The competitive effect of anti‐CypA mAb on CypA‐TβR2 interaction was detected using competitive SPR assay (G). Linearities of the CypA‐TβR2 interaction were generated from the data of the sampling points shown in (H). (I) Immunoblot analysis of the indicated proteins in A549 cells treated with eCypA (10 nM) or eCypA (10 nM) + TGF‐β pAb (50 nM) for 30 min. (J) Adherence assays of GAS in A549 cells treated with eCypA (10 nM) or eCypA (10 nM) + TGF‐β pAb (50 nM) for 12 h, and then infected with GAS (MOI = 10.0) for 2 h. Data are reported as the percentage of adherent bacteria relative to the input inoculum. (K) Immunoblot analysis of the indicated proteins in immunoprecipitated samples and WCLs of A549 cells treated with TGF‐β1 (5 ng/mL), TGF‐β1 + CypA‐His (10 nM), or TGF‐β1 + CypA‐His + anti‐CypA mAb (50 nM) for 2 h. (L and M) Interactions of TGF‐β1 with TβR2 were detected by SPR in the absence (L) or presence (M) of CypA. Data are presented as the means ± SDs; ** p < 0.01, *** p < 0.001 and ns = not significant; unpaired two‐tailed Student's t test for (D); one‐way ANOVA followed by Dunnett's test for (A) and (J).

Article Snippet: Moreover, siRNAs targeting TGFBR2 and ACE2 were purchased from Santa Cruz Biotechnology.

Techniques: Binding Assay, Analysis, Transfection, Western Blot, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Negative Control, Blocking Assay, SPR Assay, Generated, Sampling, Infection, Bacteria, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Immune and inflammatory features associated with pyroptosis- and myocardial fibrosis-related genes in HFpEF: a machine learning study

doi: 10.3389/fimmu.2026.1925067

Figure Lengend Snippet: Establishment of the HFD/L-NAME-induced HFpEF mouse model and in vivo validation of hub PMFRDEGs. Unless otherwise stated, n = 6 animals per group, and each plotted biological replicate represents one animal. (A) Body-weight trajectory (g), analyzed by two-way repeated-measures ANOVA with time and group as factors and animal as the repeated factor; sphericity was not assumed, the Geisser–Greenhouse correction was applied, and control-versus-HFpEF comparisons at individual time points were adjusted using the Šídák procedure. (B–D) Systolic, diastolic, and mean arterial pressure (mmHg), respectively. (E) Treadmill running distance (m). (F) Lung wet-to-dry weight ratio (dimensionless). (G) Representative M-mode echocardiographic images. (H, I) LVEF and LVFS (%). (J) Representative pulsed-wave and tissue-Doppler images. (K) E/e′ ratio (dimensionless). (L) Representative H&E- and Masson-stained myocardial sections. (M) Cardiomyocyte cross-sectional area (μm²). (N) Fibrotic area (%). (O–Q) Relative myocardial mRNA abundance of Ptpn11 , Pparg , and Ace2 , respectively, expressed as 2 −ΔΔ Ct relative values. (R) Representative Western blots of PTPN11, PPARG, and ACE2, with GAPDH as the loading control; full uncropped blots are provided in . (S–U) GAPDH-adjusted densitometric ratios for PTPN11, PPARG, and ACE2, respectively (dimensionless). (V) Representative immunohistochemical staining for PTPN11, PPARG, and ACE2. (W–Y) Animal-level IOD values for PTPN11, PPARG, and ACE2, respectively (a.u.). Two-sided unpaired Student’s t-tests were used for panels (B, C, E, F, H, K, P, S–U, W, X) two-sided Welch’s t-tests were used for panels (I, N, O, Y) and two-sided Mann–Whitney U tests were used for panels (D, M) , and (Q) No multiplicity adjustment was applied to these single-time-point two-group comparisons; their P values are nominal and two-sided. For Ace2 mRNA in panel (Q) n = 5 in the control group and n = 6 in the HFpEF group because one control measurement was not quantifiable and the missing value was not imputed. For panel (A) asterisks denote Šídák-adjusted P values; for all other quantitative panels, asterisks denote nominal two-sided P values. ns, P ≥ 0.05; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001.

Article Snippet: The membranes were incubated at 4 °C overnight with primary antibodies against GAPDH (1:100,000; 60004-1-Ig; Proteintech), PTPN11 (1:2,000; 66795-1-Ig; Proteintech), PPARG (1:10,000; 81490-5-RR; Proteintech), and ACE2 (1:4,000; 66699-1-Ig; Proteintech).

Techniques: In Vivo, Biomarker Discovery, Control, Tissue, Staining, Western Blot, Immunohistochemical staining, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Immune and inflammatory features associated with pyroptosis- and myocardial fibrosis-related genes in HFpEF: a machine learning study

doi: 10.3389/fimmu.2026.1925067

Figure Lengend Snippet: Internal evaluation of the fixed three-gene Firth classifier in the merged murine discovery cohort (control, n = 20; HFpEF, n = 25; total n = 45). (A) Apparent ROC curve of the Firth model (AUC = 1.0000); 2,000 stratified bootstrap resamples yielded an optimism-corrected AUC of 0.9971 (95% bootstrap percentile interval, 0.9780–1.0000). (B) Firth-based calibration analysis; the optimism-corrected calibration intercept was 0.2428 (95% bootstrap percentile interval, −0.7402–1.2763), and the calibration slope was 1.6710 (0.8315–3.1674). (C) In-sample decision curve analysis based on Firth-model predictions. (D) Nomogram reconstructed from the fitted Firth coefficients. Repeated nested cross-validation results are reported separately in the Results and are not represented by panel (A, E) Normalized log2(FPKM + 1) expression of Pparg , Ptpn11 , and Ace2 in HFpEF and control samples; groups were compared using two-sided Wilcoxon rank-sum tests, with no multiplicity adjustment. (F–H) Individual ROC curves for Pparg , Ptpn11 , and Ace2 . (I) GO semantic-similarity analysis. In panel E , asterisks denote nominal two-sided P values: ns, P ≥ 0.05; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. Asterisk notation is not used for panels (A–D) or (F–I) .

Article Snippet: The membranes were incubated at 4 °C overnight with primary antibodies against GAPDH (1:100,000; 60004-1-Ig; Proteintech), PTPN11 (1:2,000; 66795-1-Ig; Proteintech), PPARG (1:10,000; 81490-5-RR; Proteintech), and ACE2 (1:4,000; 66699-1-Ig; Proteintech).

Techniques: Control, Analysis, Biomarker Discovery, Expressing

Journal: Frontiers in Immunology

Article Title: Immune and inflammatory features associated with pyroptosis- and myocardial fibrosis-related genes in HFpEF: a machine learning study

doi: 10.3389/fimmu.2026.1925067

Figure Lengend Snippet: Exploratory replication of the three-gene panel in the human HFpEF_2020 left-ventricular dataset (control, n = 45; HFpEF, n = 41; total n = 86). The logistic model was re-estimated within HFpEF_2020 rather than evaluated using locked coefficients from the murine discovery cohort. (A) ROC curve of the refitted three-gene model. (B, C) Calibration and decision-curve analyses of the refitted model. (D) Dataset-specific nomogram generated from the HFpEF_2020 coefficients. (E) Normalized log2(FPKM + 1) expression of PPARG , PTPN11 , and ACE2 in HFpEF and control samples; groups were compared using two-sided Wilcoxon rank-sum tests, without multiplicity adjustment. (F–H) Individual ROC curves for PPARG , PTPN11 , and ACE2 . In panel (E) , asterisks denote nominal two-sided P values: ns, P ≥ 0.05; ∗ P < 0.05; ∗∗ P < 0.01; ∗∗∗ P < 0.001; ∗∗∗∗ P < 0.0001. Asterisk notation is not used for panels (A–D) or (F–H) .

Article Snippet: The membranes were incubated at 4 °C overnight with primary antibodies against GAPDH (1:100,000; 60004-1-Ig; Proteintech), PTPN11 (1:2,000; 66795-1-Ig; Proteintech), PPARG (1:10,000; 81490-5-RR; Proteintech), and ACE2 (1:4,000; 66699-1-Ig; Proteintech).

Techniques: Control, Generated, Expressing

Journal: PLOS Biology

Article Title: Targeting of conserved spike epitopes enables broad antibody neutralisation of diverse bat sarbecoviruses

doi: 10.1371/journal.pbio.3003882

Figure Lengend Snippet: A) Binding of IgG and Fab to recombinant RBDs of Sarbecoviruses from different clades by ELISA including SARS-CoV-2 (clade 1b), WIV-1 (clade 1a), Rs4321 (clade 1a), RacCS203 (clade 2), Rc-o319 (clade 5), and RhGB07 (clade 3). Experiments were run in duplicate and repeated twice. A representative assay is shown. Fab binding is shown with the dotted line and IgG binding with the full line. B) Neutralising activity of IgGs against SARS-CoV-2 and variants of concern as well as SARS-CoV-1 and WIV-1. C) IC 50 values for neutralisation against SARS-CoV-2 variants of concern and other sarbecoviruses that utilise human-ACE2. For A–C, the mean value is plotted with error bars showing ± standard deviation (s.d.). D) Ability of IgGs to inhibit the interaction between cell surface human ACE2 and soluble SARS-CoV-2 and WIV-1 RBDs. IgGs were pre-incubated with fluorescently labelled RBD before addition to HeLa-ACE2 cells. The percentage reduction in mean fluorescence intensity is reported. Experiments were performed in duplicate. E) Superimposed 3D structures of the V1WT_06 (dark blue), V1WT_41 (purple), and VA14_26 (red) around WIV-1 RBD (light blue, from PDB 8WLZ). The data underlying this Figure can be found in .

Article Snippet: HEK293T cells were transfected with either Human, Halcyon or Japanese horseshoe bat ACE2 using PEI Max (1 mg/mL, Polysciences) at 3:1, 6:1, and 2:1 ratio (PEI Max:DNA), respectively.

Techniques: Binding Assay, Recombinant, Enzyme-linked Immunosorbent Assay, Activity Assay, Standard Deviation, Incubation, Fluorescence

Journal: PLOS Biology

Article Title: Targeting of conserved spike epitopes enables broad antibody neutralisation of diverse bat sarbecoviruses

doi: 10.1371/journal.pbio.3003882

Figure Lengend Snippet: A) A closeup view of the superimposed structure of V1WT_06 bound to WIV-1 RBD on a trimeric spike (from PDB: 8WLZ). A clash with NTD of chain B is observed when V1WT_06 Fab-RBD complex is aligned onto RBD in the up conformation (chain A) in trimeric WIV-1 spike protein. Notably, the binding site is inaccessible in the RBD-down conformation. B) V1WT_06 heavy chain only interaction with WIV-1 RBD (grey). Heavy and light chains are shown with the dark and light blue ribbons, respectively. C) V1WT_06 epitope footprint overlayed with epitope footprints of other site V neutralising antibodies including S2H97, CC25.4, 5817, R1-32, and E465 patch [ , – , ]. SARS-CoV-2 RBD (from PDB: 6m0j) is coloured based on the amino acid conservation across 87 bat sarbecoviruses [ , , , ]. D) Sequence alignment of a panel of bat sarbecovirus RBDs. Contact residues of V1WT_06 mAb on the WIV-1-RBD are indicated with black dots. The colouring scheme describes the percent identity (PID) between the sequences as implemented in Jalview 2.11.4.0. E) Comparison in neutralisation potency of V1WT_06 Fab and IgG against SARS-CoV-2 (D614G), SARS-CoV-1, and WIV-1. Mean values are plotted with error bars showing ± standard deviation (s.d.). The data underlying this Figure can be found in .

Article Snippet: HEK293T cells were transfected with either Human, Halcyon or Japanese horseshoe bat ACE2 using PEI Max (1 mg/mL, Polysciences) at 3:1, 6:1, and 2:1 ratio (PEI Max:DNA), respectively.

Techniques: Binding Assay, Sequencing, Comparison, Standard Deviation

Journal: PLOS Biology

Article Title: Targeting of conserved spike epitopes enables broad antibody neutralisation of diverse bat sarbecoviruses

doi: 10.1371/journal.pbio.3003882

Figure Lengend Snippet: A) VA14_26 and V1WT_41 antibody footprints overlayed with the epitope footprints of other RBD class 4 neutralising antibodies including CR3022, 10−40, ADI-62113 [ – ]. SARS-CoV-2 RBD (from PDB: 6m0j) is coloured based on the amino acid conservation across 87 bat sarbecoviruses [ , , , ]. B) Superimposed 3D structure depicting the binding of VA14_26 (Red) to WIV-1 RBD (blue) in relation to ACE2 (orange, from PDB: 8WLZ) and CR3022 (pink, from PDB: 6W41). C) Superimposed 3D structure depicting the binding of V1WT_41 (green) to WIV-1 RBD (blue) in relation to ACE2 (orange, from PDB: 8WLZ). D) Sequence alignment of a panel of bat sarbecovirus RBDs. Contact residues for VA14_26 and V1WT_41 are indicated by red and pink dots and outline, respectively. Shared contact residues are shown with yellow dots and outline. E) Sequence alignment of a panel of SARS-CoV-2 variant RBDs. Contact residues for VA14_26 and V1WT_41 are indicated by red and pink dots and outline, respectively. Shared contact residues are shown with yellow dots and outline. The colouring scheme describes the percent identity (PID) between the sequences as implemented in Jalview 2.11.4.0. F) A close-up view of YYDRSG motif in V1WT_41 HCDR3 (green) in contact with WIV-1 RBD (grey), shown in stick representation, renumbered based on alignment to SARS-CoV-2 [ , ]. Presence of G109 breaks the H-bonding pattern in the beta hairpin to create a G1 β-bulge. This further enables D106 side chain to make hydrogen bond with the amides in the protein backbone, thereby stabilising the local structure. Combined this accentuates a right-handed twist found in G1 β-bulge . Twisting of the β-hairpin is thought to contribute towards stabilisation of RBD-Fab interaction in several other public mAbs with YYDRxG motif [ , ].

Article Snippet: HEK293T cells were transfected with either Human, Halcyon or Japanese horseshoe bat ACE2 using PEI Max (1 mg/mL, Polysciences) at 3:1, 6:1, and 2:1 ratio (PEI Max:DNA), respectively.

Techniques: Binding Assay, Sequencing, Variant Assay

Journal: PLOS Biology

Article Title: Targeting of conserved spike epitopes enables broad antibody neutralisation of diverse bat sarbecoviruses

doi: 10.1371/journal.pbio.3003882

Figure Lengend Snippet: Neutralisation of sarbecoviruses through entry via different host ACE2s. Neutralisation of A) Rc-o319 and B) RhGB07 was measured against entry via Rh. cornutus ACE2 and Rh. alcyone ACE2, respectively, which were transiently expressed in HEK293T/17 cells. C) Neutralisation of WIV-1 was compared between entry via hu-ACE2 (dotted line and closed symbols) and Rh. alcyone ACE2 (full line and closed symbols). Neutralisation was measured using HEK 293T cells transiently expressing either Human ACE2 or Rh. alcyone ACE2. For A-C, mean values are plotted with error bars showing ± standard deviation (s.d.). D) Ability of IgGs to inhibit the interaction between cell surface ACE2 (including hu-ACE2, Rh. cornutus ACE2 and Rh. alcyone ACE2) and soluble RBD (including SARS-CoV-2, WIV-1, Rs4231, RhBG07, and Rc-o319 RBDs). IgGs were pre-incubated with fluorescently labelled RBD before addition to ACE2 expressing cells. N.B = No binding of IgG to the RBD. The percentage reduction in mean fluorescence intensity is reported. Experiments were performed in duplicate. E) Antibody dependant cellular cytotoxicity (ADCC) of IgGs measured through the surface expression of CD107a on activated NK cells (NK-92 cells expressing CD16) . HIV-1 specific neutralising antibody, PGT128 is used as a negative control . The data underlying this Figure can be found in .

Article Snippet: HEK293T cells were transfected with either Human, Halcyon or Japanese horseshoe bat ACE2 using PEI Max (1 mg/mL, Polysciences) at 3:1, 6:1, and 2:1 ratio (PEI Max:DNA), respectively.

Techniques: Expressing, Standard Deviation, Incubation, Binding Assay, Fluorescence, Negative Control