Journal: medRxiv

Article Title: Identification of CAA as highly specific and sensitive antibody target for acute schistosomiasis diagnostics

doi: 10.1101/2023.03.07.23286891

Figure Lengend Snippet: Average SmCA (circles) and SmSEA (triangles) specific A) IgM and B) IgG antibody levels elicited in response to controlled human schistosome infection (CSI) with male-only cercariae (dose 10/20/30 cercariae, n=17 individuals) over time. No parasite eggs were produced in this single-sex CSI model and hence all antibodies recognizing SmSEA are binding to shared, predominantly glycan, epitopes present in both SmCA and SmSEA. Tx indicates treatment with 40mg/kg praziquantel at twelve weeks post infection. Heatmaps showing levels (log2 MFI+1) of IgG1 and IgG2 specific for C) SmCA and D) SmSEA mock (M) and periodate treated (P) antigen for six individuals infected with 10 or 30 cercariae (10 cercs: IX, X, XI, 30 cercs: XII, XIII, XIV) at an early phase (weeks 4, 6, 8) and later phase (weeks 12, 16, 20) post infection. Note the highly similar pattern for SmCA and SmSEA, which is due to epitopes shared between SmCA and SmSEA (male only infection, no eggs present). No anti-SmCA/SmSEA IgG3 or IgG4 was detectable for IX, X, XI, not measured for XII, XIII, XIV. Any binding below log2 (MFI+1) = 8 is negligible. X = no data available. E) IgM and F) IgG antibodies recognising SmCA (blue circles) and SmSEA (purple triangles) are present in samples from Dutch blood donors (donor n=56), from Dutch pre-travel samples (PTSP n=131), from LUMC biobank samples with S. stercoralis infection ( S. ster . n=25), and from samples with Trichiura trichiura, Ascaris sp., and/or hookworm infections from Flores Island, Indonesia which is also non-endemic for schistosomes (STH IgM n=87, IgG n=97). For each individual in each sample set paired data for mock treated (M, left) and periodate treated (P, right) soluble antigen is shown. A decrease in MFI for the vast majority of antibody binding in mock compared to periodate treated antigen illustrates, that many antibodies measured are recognising glycan elements present within the crude antigen mixes. Such shared glycan epitopes pose a challenge for specificity and hence accuracy in diagnostic immunoassays. See Suppl. for specificity calculations based on the arbitrary cut-off (MFI=1×10 4 ) shown as a dotted line on E) and F). All data shown as mean fluorescence intensity (MFI) measured on microarray. SD = standard deviation.

Article Snippet: For IgG subclass experiments 1:400 mouse anti-human (Southern Biotech) IgG1 Hinge-AF555 (#9052-32), IgG2 Fc-AF555 (#9070-32), and IgG3 Hinge-AF647 (#9210-31) were used and IgG4 pFc’-AF647 (#9190-31) in 1:250.

Techniques: Infection, Produced, Binding Assay, Glycoproteomics, Diagnostic Assay, Fluorescence, Microarray, Standard Deviation

Journal: medRxiv

Article Title: Identification of CAA as highly specific and sensitive antibody target for acute schistosomiasis diagnostics

doi: 10.1101/2023.03.07.23286891

Figure Lengend Snippet: Average anti-circulating anodic antigen (CAA, rhombes) and anti-circulating cathodic antigen (CCA, circles) specific A) IgM and B) IgG antibody levels induced in CSI with male-only cercariae (dose 10/20/30 cercariae, n=17 individuals) over time at baseline (0) and at one, two, three, four, five, six, seven, eight, ten, 12, 14, 16, 18, 20 and 52 weeks post-infection. For B) anti-CAA IgG responses are split in two groups depicting individuals responding early (≤ week 4, rhombes) and late (≥ week 7, stars). The early responder group consist of 30 (n=3) and 20 (n=3) exposure group individuals, whereas the late responder grouping consist of 20 (n=8) and 10 (n=3) exposure group individuals. Data shown as mean fluorescence intensity (MFI) with standard deviation (SD) measured on microarray. C) Heatmap showing anti-CAA and D) anti-CCA IgG1 and IgG2 levels (log2 MFI+1) for the same six individuals as (10 cercs: IX, X, XI, 30 cercs: XII, XIII, XIV) at an early phase (weeks 4, 6, 8) and later phase (weeks 12, 16, 20) post infection. No anti-CAA/CCA IgG3 or IgG4 was detectable for IX, X, XI, not measured for XII, XIII, XIV. Any binding below log2 (MFI+1) = 8 is negligible. X = no data available.

Article Snippet: For IgG subclass experiments 1:400 mouse anti-human (Southern Biotech) IgG1 Hinge-AF555 (#9052-32), IgG2 Fc-AF555 (#9070-32), and IgG3 Hinge-AF647 (#9210-31) were used and IgG4 pFc’-AF647 (#9190-31) in 1:250.

Techniques: Infection, Fluorescence, Standard Deviation, Microarray, Binding Assay

Journal: medRxiv

Article Title: Identification of CAA as highly specific and sensitive antibody target for acute schistosomiasis diagnostics

doi: 10.1101/2023.03.07.23286891

Figure Lengend Snippet: IgM (A-D) and IgG (E-H) responses to CAA, CCA, S. mansoni CA (SmCA), S. haematobium CA (ShCA), S. mansoni SEA (SmSEA), and S. haematobium SEA (ShSEA) for schistosomiasis infection naïve individuals are shown as dotblots for: Dutch blood donors (A and E, n=56 NLD donor)), Dutch pre-travel samples (B and F, n=IgM 130/IgG 131 PTSP), S. stercoralis infected LUMC biobank samples (C and H, n=25), and soil-transmitted helminth ( Trichuris trichiura, Ascaris sp., and/or hookworm) infected individuals from Pulau Flores, Indonesia (D and H, n=IgM 87/IgG 97 STH). All data shown as mean fluorescence intensity (MFI) measured on microarray with medians indicated as horizontal bars. See for specificity calculations based on the arbitrary cut-off (MFI=10000) shown as a dotted lines on graphs (A-H).

Article Snippet: For IgG subclass experiments 1:400 mouse anti-human (Southern Biotech) IgG1 Hinge-AF555 (#9052-32), IgG2 Fc-AF555 (#9070-32), and IgG3 Hinge-AF647 (#9210-31) were used and IgG4 pFc’-AF647 (#9190-31) in 1:250.

Techniques: Infection, Fluorescence, Microarray

Journal: medRxiv

Article Title: Identification of CAA as highly specific and sensitive antibody target for acute schistosomiasis diagnostics

doi: 10.1101/2023.03.07.23286891

Figure Lengend Snippet: IgM (A) and IgG (B) responses to CAA and CCA in six young male Spanish travellers exposed to S. mansoni clade parasite in Chicamba Lake, Mozambique in 2019 through a single swimming activity . Individuals shown with dashed lines are paired for A and B to highlight time dynamics for individuals without consistently high antibody titer. These two individuals show differences in timing of isotype switch. One individual was the first to present with symptoms at the travel clinic at day 39 after infection (low antibodies) and with high IgM and IgG at 44 days post infection and declining antibody levels from ∼20 weeks post infection. The second has lower IgM from first measurement at ∼7 weeks up to ∼20 weeks post exposure, where isotype switch occurs and CAA specific IgG is induced. C) Anti-CAA and anti-CCA for paired pre- and post-travel samples from nine adult Dutch individuals all identified as schistosomiasis IgM positive but only three IgG positive in original study . These nine individuals were exposed to Schistosoma sp. through swimming in Lake Malawi and/or Lake Victoria at separate event(s) during travel of varying lengths and measured within three months after returning to The Netherlands. All post-travel samples from individuals with no schistosome exposure were confirmed to still be negative for anti-CAA and anti-CCA (PTSP pre-travel shown in ) shows anti-CAA responses for 34 Belgian individuals travelling as part of two separate groups of families but exposed at same location through swimming and rafting in uMhkunyane river, Northern Kwa-Zulu Natal, South Africa in 2016/17 to S. haematobium clade parasites ( S. haematobium x S. matheii hybrid and/or co-infection(s)) within a two week interval ( , ), no data for anti-CCA antibodies available for this sample set due to limited sample volume. Data shown as mean fluorescence intensity (MFI) measured on microarray.

Article Snippet: For IgG subclass experiments 1:400 mouse anti-human (Southern Biotech) IgG1 Hinge-AF555 (#9052-32), IgG2 Fc-AF555 (#9070-32), and IgG3 Hinge-AF647 (#9210-31) were used and IgG4 pFc’-AF647 (#9190-31) in 1:250.

Techniques: Activity Assay, Infection, Northern Blot, Fluorescence, Microarray

Journal: EBioMedicine

Article Title: A liquid biopsy for detecting circulating mesothelial precursor cells: A new biomarker for diagnosis and prognosis in mesothelioma

doi: 10.1016/j.ebiom.2020.103031

Figure Lengend Snippet: Overview of mesothelial precursor cells (MPC) analysis using a microfluidic approach. (a) Sample preparation for MPC analysis is quick and simple. MSLN+ cells are tagged with biotinylated anti-MSLN antibody and further conjugated with anti-biotin magnetic nanoparticle (MNP) complex. Magnetically labeled samples are loaded into the microfluidic chip at 500µL/h with external magnets placed above and below the device to facilitate magnetic trapping of targeted cells. (b) Multiple populations of MPC exist in circulation and are subcategorized based on varying combinations of CD34 and/or CD90 in conjunction with mesothelin (MSLN). Image created with BioRender. (c) Overview of MesoFind microfluidic chip for MPC isolation. The microfluidic device contains 8 different zones with a sequential increase in height from 50µm to 400µm (inlet-to-outlet) and features varying average linear flow velocities (1.0x, 0.50x, 0.33x, 0.25x, 0.20x, 0.17x, 0.14x, 0.13x). Cells magnetically labeled with mesothelin are captured in different zones based on the relative amount of magnetic content. Cells with higher MSLN expression (high magnetic content) are captured in earlier zones whereas cells with less MSLN (low magnetic content) are captured in later zones. Captured cells are fluorescently labeled with CD34 and CD90 antibodies to identify and isolate different MPC subpopulations. (d) MSLN expression profiles are generated for each MPC subpopulation based on the captured location on the chip. Each MPC subtypes are predicted to exhibit different clinical characteristics in patients and their expression of MSLN correlates to the severity of the conditions.

Article Snippet: The captured cells were then immunostained with 200µL of an antibody cocktail containing either 2mg/mL of anti-CD34 AF488 (bs-8996R-A488, Bioss Antibodies, USA), 2mg/mL of anti-CD90 AF555 (bs-0778R-A555, Bioss Antibodies, USA), and 0.6 mg/mL anti-CD45 APC (17-0451-83, Invitrogen, USA) for mice-related samples or 1mg/mL of anti-CD34 AF488 (ab195013, Abcam, Canada), 0.5mg/mL of anti-CD90 AF555 (bs-10430R-A555, Bioss Antibodies, USA), and 1.2% v/v of anti-CD45 APC (555485, BD Biosciences, USA) for human-related samples.

Techniques: Sample Prep, Labeling, Isolation, Expressing, Generated

Journal: EBioMedicine

Article Title: A liquid biopsy for detecting circulating mesothelial precursor cells: A new biomarker for diagnosis and prognosis in mesothelioma

doi: 10.1016/j.ebiom.2020.103031

Figure Lengend Snippet: Validation of the microfluidic approach for MPC analysis. (a) Captured efficiency of mice mesothelioma cell line (RN5) and human mesothelioma cell lines (H2052 and H2452) spiked in healthy blood using MesoFind microfluidic chips. (b) Identification of different MPC and leukocytes from blood using fluorescence microscopy after cell entrapment using MesoFind devices. Three differing MPC subtypes (top three) shown here contains different combinations of CD34 and/or CD90 expression and were negative for CD45. Leukocytes shown express CD34- CD90- CD45+ but were generally identified as CD45+ and can have varying expressions of CD34 and CD90 (not shown). All scale bars are 20 um. (c) Schematic of mesothelioma mice model analysis. RN5 mesothelioma cells were injected intraperitoneally (i.p.) and mice were sacrificed between 1 to 6 weeks post-injection. Control mice were injected with saline. The lavage and blood collected from the mice were analyzed using flow cytometry or MesoFind microfluidic analysis followed by fluorescence microscopy for the identification of different MPC populations. Images were created with BioRender. (d) Total MSLN+ CD34+ CD90+ MPC captured from the lavage collected from mice using MesoFind analysis. A general increase in total counts observed from 0 to 6 weeks post-injection. (e) Flow cytometry of MSLN+ CD34+ CD90+ MPC in lavage samples. No significant change was observed after 4-weeks post-injection. No MPC were detected in blood samples using flow cytometric analysis (not shown). (f) Total MSLN+ CD34+ CD90+ MPC captured from the blood of mice using MesoFind analysis. A general increase in total counts observed from 0 to 6 weeks post-injection. (g) Mesothelin expression indices (EI) for MSLN+ CD34+ CD90+ MPC in lavage and blood samples from 0 to 6 weeks post-injection using MesoFind analysis. (h) Flow cytometric gating for MSLN+ cells and CD34+ CD90+ cells in naive mice and (i) 4 weeks RN5 post-injected mice. All analysis was performed in triplicates (n=3). Error bars for Fig. 2 d,f, and g represent standard error of mean. All reported values were compared to values at 0 week (d,e,f) or naïve mice (g) using unpaired t-test, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The captured cells were then immunostained with 200µL of an antibody cocktail containing either 2mg/mL of anti-CD34 AF488 (bs-8996R-A488, Bioss Antibodies, USA), 2mg/mL of anti-CD90 AF555 (bs-0778R-A555, Bioss Antibodies, USA), and 0.6 mg/mL anti-CD45 APC (17-0451-83, Invitrogen, USA) for mice-related samples or 1mg/mL of anti-CD34 AF488 (ab195013, Abcam, Canada), 0.5mg/mL of anti-CD90 AF555 (bs-10430R-A555, Bioss Antibodies, USA), and 1.2% v/v of anti-CD45 APC (555485, BD Biosciences, USA) for human-related samples.

Techniques: Fluorescence, Microscopy, Expressing, Injection, Flow Cytometry

Journal: EBioMedicine

Article Title: A liquid biopsy for detecting circulating mesothelial precursor cells: A new biomarker for diagnosis and prognosis in mesothelioma

doi: 10.1016/j.ebiom.2020.103031

Figure Lengend Snippet: Clinical utility of analyzing various MPC subpopulations using MesoFind device. (a) Total blood counts for MSLN+ CD34+ CD90+ MPC from healthy donor (HD, n=10), asbestos-exposed individuals (ASB, n=23), and malignant pleural mesothelioma patients (MPM, n=23) using MesoFind devices. (b) Comparison of mesothelin expression indices between varying subpopulations of MPC (CD90+ CD34-, CD90- CD34+, CD90+ CD34+, CD90+, CD34+) in HD, ASB, and MPM individuals. All MPC were MSLN+ and CD45-. (c-h) MSLN expression indices (EI) for most representative MPC subtypes in each clinical evaluation. Overall, higher MSLN expression indices corresponded to higher severity in clinical tests. EI cut-off values between each group were determined and the clinical sensitivity and specificity were reported (right). (c) EI of MSLN+ CD90- CD34+ CD45- MPC in ASB (n=23) and MPM (n=23). Cut-off EI=500. (d) EI of MSLN+ CD90+ CD34- CD45- MPC in HD (n=10) and ASB (n=23). Cut-off EI=200. (e) EI of MSLN+ CD90+ CD45- MPC were compared between epithelioid (n=16) and biphasic (n=5) cancer subtypes. Cut-off EI = 2650. (f) EI of MSLN+ CD34+ CD45- MPC in patients with low (<6) (n=4) and high (>6) (n=4) SUV scores through PET scans were compared. Cut-off EI=6700. (g) EI of MSLN+ CD90+ CD34- CD45- MPC for patients with unresectable tumors (n=19) and resectable tumors (n=4) were compared. Cut-off EI=200. (h) Presence (n=3) and absence (n=13) of lymph node metastasis was compared through MSLN+ CD34+ CD45- MPC. Cut-off EI=24500. Statistics are performed with unpaired t-tests, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The captured cells were then immunostained with 200µL of an antibody cocktail containing either 2mg/mL of anti-CD34 AF488 (bs-8996R-A488, Bioss Antibodies, USA), 2mg/mL of anti-CD90 AF555 (bs-0778R-A555, Bioss Antibodies, USA), and 0.6 mg/mL anti-CD45 APC (17-0451-83, Invitrogen, USA) for mice-related samples or 1mg/mL of anti-CD34 AF488 (ab195013, Abcam, Canada), 0.5mg/mL of anti-CD90 AF555 (bs-10430R-A555, Bioss Antibodies, USA), and 1.2% v/v of anti-CD45 APC (555485, BD Biosciences, USA) for human-related samples.

Techniques: Expressing

Journal: Cell Stem Cell

Article Title: Defining the Identity and Dynamics of Adult Gastric Isthmus Stem Cells

doi: 10.1016/j.stem.2019.07.008

Figure Lengend Snippet: Rapidly Cycling Isthmus Progenitors Can Maintain Long-Term Self-Renewal Potential (A) Schematic showing the fluorescence signal associated with each phase of the cell cycle in the Rosa26-Fucci2a mouse. (B) Representative confocal images of stomach corpus gland sections (150 μm) of Rosa26-Fucci2a mice. Red, hCdt1-mCherry(30/120-G1); green, hGem(1/110-S/G2/M); grey, β-catenin (I), Ki67 (II), HKATPase (III). Scale bars: 50 μm. (C) Venn diagram showing the overlap between genes more highly expressed in isolated proliferative isthmus cells and genes downregulated following 5-FU treatment, taken to be prospective candidate markers for proliferating isthmus cells. (D) Immunohistochemical staining for Ki67, counterstained with Mayer’s hematoxylin (leftmost panel). In situ hybridization with Stmn1 anti-sense (middle panel) and Stmn1 sense negative control (rightmost panel) probes in mouse stomach corpus sections are shown. Stmn1 mRNA was restricted to the isthmus region delineated by Ki67 staining. Scale bars: 25 μm. (E) Schematic of the genetic strategy to trace Stmn1+ cell-derived progeny in the corpus gland. (F) Representative confocal images of the stomach corpus of Stmn1-CreERT2 mice. Red, Ki67; green, EGFP. Scale bars: 50 μm. (G) Representative confocal images of 150-μm-thick sections of the stomach corpus of Stmn1-CreERT2;R26R-tdTomato mice at 2 days, 2 weeks, 1 month, 3 months, and 6 months post-injection of 0.5–1 mg tamoxifen/20 g mouse body weight. Labeling of clones in the isthmus region with tdTomato can be detected as early as 2 days post-injection. Red, tdTomato. Scale bars: 50 μm. (H) Representative confocal images of the mouse stomach corpus glands of Stmn1-CreERT2;R26R-tdTomato mice at 5 months post-labeling. Red, tdTomato for labeled cells; grey, Atp4b for parietal cells; green, Muc5ac for pit cells and GS-II for neck cells. Scale bars: 50 μm. See Figure S6 and and .

Article Snippet: Sections were incubated in blocking solution (2% donkey or goat serum, 5% DMSO and 0.5% Triton X-100 in PBS) at room temperature for 2 h. The following primary antibodies were used: chicken anti-EGFP (1:500; Abcam, ab13970), rabbit anti-Ki67 (1:250; A. Menarini, MP-325-CRM1), Alexa 647-conjugated mouse anti-β-catenin (1:200; Cell Signaling Technology, 4627S), rabbit anti-H+/K+ ATPase β Antibody (D-18) (1:500; Santa Cruz, sc-84304) and Alexa 555/647-conjugated rabbit anti-ATP4b (1:200; Bioss, bs-2433R-AF555, bs-2433R-AF647).

Techniques: Fluorescence, Isolation, Immunohistochemical staining, Staining, In Situ Hybridization, Negative Control, Derivative Assay, Injection, Labeling, Clone Assay

Journal: Cell Stem Cell

Article Title: Defining the Identity and Dynamics of Adult Gastric Isthmus Stem Cells

doi: 10.1016/j.stem.2019.07.008

Figure Lengend Snippet:

Article Snippet: Sections were incubated in blocking solution (2% donkey or goat serum, 5% DMSO and 0.5% Triton X-100 in PBS) at room temperature for 2 h. The following primary antibodies were used: chicken anti-EGFP (1:500; Abcam, ab13970), rabbit anti-Ki67 (1:250; A. Menarini, MP-325-CRM1), Alexa 647-conjugated mouse anti-β-catenin (1:200; Cell Signaling Technology, 4627S), rabbit anti-H+/K+ ATPase β Antibody (D-18) (1:500; Santa Cruz, sc-84304) and Alexa 555/647-conjugated rabbit anti-ATP4b (1:200; Bioss, bs-2433R-AF555, bs-2433R-AF647).

Techniques: Recombinant, Blocking Assay, Labeling, Multiplex Assay, Isolation, Software

Journal: Journal of ethnopharmacology

Article Title: Efficacy and potential therapeutic mechanism of Weiwei decoction on Spasmolytic polypeptide-expressing metaplasia in Helicobacter pylori-infected and Atp4a-knockout mice.

doi: 10.1016/j.jep.2023.117062

Figure Lengend Snippet: Fig. 3. WWD treatment ameliorates SPEM in both Helicobacter pylori-infected and Atp4a-/- mice. (A) Expression of clusterin (CLU, AF555) and Griffonia simplicifolia lectin II (GSII, AF488) in H. pylori-infected mice was determined by immunofluorescence with a fluorescence microscope. Nuclei were stained with Hoechst 33342, scale bar = 50 μm. Analysis of CLU+GSII+ (B), GSII (C), and CLU (D) signal area in H. pylori mice normalized by the Control group, n = 3–4. Data are presented as mean ± SEM. ###p < 0.001 vs. control group, *p < 0.05,**p < 0.01, ***p < 0.01, vs. H. pylori group. (E) The expression of CLU(AF555) and GSII(AF488) in Atp4a-/-

Article Snippet: After washing, the samples were incubated for 2 h at room temperature with goat anti-rabbit IgG AF555 (1:200, 1036-30, Southern Biotech) or goat anti-mouse IgG AF555 (1:200, 1036-32, Southern Biotech, USA) secondary antibody diluted in PBST (PBS with 0.05% Tween-20).

Techniques: Infection, Expressing, Immunofluorescence, Fluorescence, Microscopy, Staining, Control

Journal: Journal of ethnopharmacology

Article Title: Efficacy and potential therapeutic mechanism of Weiwei decoction on Spasmolytic polypeptide-expressing metaplasia in Helicobacter pylori-infected and Atp4a-knockout mice.

doi: 10.1016/j.jep.2023.117062

Figure Lengend Snippet: Fig. 7. WWD treatment partially restores GIF expression in Atp4a-/- mice. (A) Expression of gastric intrinsic factor (GIF, AF555, red) and Griffonia simplicifolia lectin II (GSII, AF488, green) in Atp4a-/- mice was determined by immunofluorescence with laser confocal microscope. Nuclei were stained with Hoechst 33342 (blue). Magnification: 100 × and 200 × , scale bar = 50 μm. (B) Analysis of GIF signal area in each group of Atp4a-/- mice, n = 3, two images of each sample were used for statistical analysis. Data are presented as mean ± SEM. #p < 0.05, vs. Atp4a+/+ group. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After washing, the samples were incubated for 2 h at room temperature with goat anti-rabbit IgG AF555 (1:200, 1036-30, Southern Biotech) or goat anti-mouse IgG AF555 (1:200, 1036-32, Southern Biotech, USA) secondary antibody diluted in PBST (PBS with 0.05% Tween-20).

Techniques: Expressing, Immunofluorescence, Microscopy, Staining

Journal: Journal of ethnopharmacology

Article Title: Efficacy and potential therapeutic mechanism of Weiwei decoction on Spasmolytic polypeptide-expressing metaplasia in Helicobacter pylori-infected and Atp4a-knockout mice.

doi: 10.1016/j.jep.2023.117062

Figure Lengend Snippet: Fig. 10. WWD treatment decreases CD163+F4/80+ M2 macrophages in gastric mucosa in Helicobacter pylori-infected mice. (A) Expression of CD163 (AF555) and F4/ 80 (AF488) in H. pylori mice was determined by immunofluorescence with a fluorescence microscope. Nuclei were stained with Hoechst 33342. Scale bar = 50 μm. (B) Analysis of CD163+ F4/80+ signal area in each group of H. pylori mice after WWD treatment, n = 4, three images of each sample were used for statistical analysis. Relative mRNA expression levels of Tnfa (C), Il1b (D), and Il33 (E) in each group of H. pylori mice, n = 8–10. Data are presented as mean ± SEM. ##p < 0.01, ###p < 0.001 vs. Control group. **p < 0.01,***p < 0.001 vs. H. pylori group.

Article Snippet: After washing, the samples were incubated for 2 h at room temperature with goat anti-rabbit IgG AF555 (1:200, 1036-30, Southern Biotech) or goat anti-mouse IgG AF555 (1:200, 1036-32, Southern Biotech, USA) secondary antibody diluted in PBST (PBS with 0.05% Tween-20).

Techniques: Infection, Expressing, Immunofluorescence, Fluorescence, Microscopy, Staining, Control

Journal: Journal of ethnopharmacology

Article Title: Efficacy and potential therapeutic mechanism of Weiwei decoction on Spasmolytic polypeptide-expressing metaplasia in Helicobacter pylori-infected and Atp4a-knockout mice.

doi: 10.1016/j.jep.2023.117062

Figure Lengend Snippet: Fig. 3. WWD treatment ameliorates SPEM in both Helicobacter pylori-infected and Atp4a-/- mice. (A) Expression of clusterin (CLU, AF555) and Griffonia simplicifolia lectin II (GSII, AF488) in H. pylori-infected mice was determined by immunofluorescence with a fluorescence microscope. Nuclei were stained with Hoechst 33342, scale bar = 50 μm. Analysis of CLU+GSII+ (B), GSII (C), and CLU (D) signal area in H. pylori mice normalized by the Control group, n = 3–4. Data are presented as mean ± SEM. ###p < 0.001 vs. control group, *p < 0.05,**p < 0.01, ***p < 0.01, vs. H. pylori group. (E) The expression of CLU(AF555) and GSII(AF488) in Atp4a-/-

Article Snippet: After washing, the samples were incubated for 2 h at room temperature with goat anti-rabbit IgG AF555 (1:200, 1036-30, Southern Biotech) or goat anti-mouse IgG AF555 (1:200, 1036-32, Southern Biotech, USA) secondary antibody diluted in PBST (PBS with 0.05% Tween-20).

Techniques: Infection, Expressing, Immunofluorescence, Fluorescence, Microscopy, Staining, Control

Journal: Journal of ethnopharmacology

Article Title: Efficacy and potential therapeutic mechanism of Weiwei decoction on Spasmolytic polypeptide-expressing metaplasia in Helicobacter pylori-infected and Atp4a-knockout mice.

doi: 10.1016/j.jep.2023.117062

Figure Lengend Snippet: Fig. 7. WWD treatment partially restores GIF expression in Atp4a-/- mice. (A) Expression of gastric intrinsic factor (GIF, AF555, red) and Griffonia simplicifolia lectin II (GSII, AF488, green) in Atp4a-/- mice was determined by immunofluorescence with laser confocal microscope. Nuclei were stained with Hoechst 33342 (blue). Magnification: 100 × and 200 × , scale bar = 50 μm. (B) Analysis of GIF signal area in each group of Atp4a-/- mice, n = 3, two images of each sample were used for statistical analysis. Data are presented as mean ± SEM. #p < 0.05, vs. Atp4a+/+ group. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: After washing, the samples were incubated for 2 h at room temperature with goat anti-rabbit IgG AF555 (1:200, 1036-30, Southern Biotech) or goat anti-mouse IgG AF555 (1:200, 1036-32, Southern Biotech, USA) secondary antibody diluted in PBST (PBS with 0.05% Tween-20).

Techniques: Expressing, Immunofluorescence, Microscopy, Staining

Journal: Journal of ethnopharmacology

Article Title: Efficacy and potential therapeutic mechanism of Weiwei decoction on Spasmolytic polypeptide-expressing metaplasia in Helicobacter pylori-infected and Atp4a-knockout mice.

doi: 10.1016/j.jep.2023.117062

Figure Lengend Snippet: Fig. 10. WWD treatment decreases CD163+F4/80+ M2 macrophages in gastric mucosa in Helicobacter pylori-infected mice. (A) Expression of CD163 (AF555) and F4/ 80 (AF488) in H. pylori mice was determined by immunofluorescence with a fluorescence microscope. Nuclei were stained with Hoechst 33342. Scale bar = 50 μm. (B) Analysis of CD163+ F4/80+ signal area in each group of H. pylori mice after WWD treatment, n = 4, three images of each sample were used for statistical analysis. Relative mRNA expression levels of Tnfa (C), Il1b (D), and Il33 (E) in each group of H. pylori mice, n = 8–10. Data are presented as mean ± SEM. ##p < 0.01, ###p < 0.001 vs. Control group. **p < 0.01,***p < 0.001 vs. H. pylori group.

Article Snippet: After washing, the samples were incubated for 2 h at room temperature with goat anti-rabbit IgG AF555 (1:200, 1036-30, Southern Biotech) or goat anti-mouse IgG AF555 (1:200, 1036-32, Southern Biotech, USA) secondary antibody diluted in PBST (PBS with 0.05% Tween-20).

Techniques: Infection, Expressing, Immunofluorescence, Fluorescence, Microscopy, Staining, Control