Journal: Circulation

Article Title: Indices of Apoptosis Activation After Blood Cardioplegia and Cardiopulmonary Bypass

doi: 10.1161/circulationaha.105.000828

Figure Lengend Snippet: Figure 3. Histological immunostaing of myocardial samples revealed a trend toward increased translocation of mature AIF (molecular weight 57 kDa) from mitochondrial membrane to nucleus as an indicator of apoptosis induction via the caspase- independent pathway. White arrows point the increased number of AIF-positive nuclei after CBC/CPB. B indicates before CBC/ CPB; A, after CBC/CPB.

Article Snippet: Specific antibodies for Bcl-2, phospho-Bcl2Ser70 (p-Bcl2), Bad, phospho-BadSer112 (p-Bad), caspase 3, cleaved caspase 3, AIF, PARP, and cleaved PARP were used (Cell Signaling Tech.

Techniques: Translocation Assay, Molecular Weight, Membrane

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Astragaloside IV Alleviates Ulcerative Colitis Progression by Inhibiting WDR5 ‐Mediated ENO1 H3K4me3 Modification

doi: 10.1002/kjm2.70064

Figure Lengend Snippet: WDR5, a target of ASI, is overexpressed in UC models. (A) The chemical structural formula of ASI was obtained from PubChem Substance. (B) Transcriptional differences between colonic biopsies from patients with UC and non‐inflammatory controls in the GSE38713 dataset. (C) The intersection of differentially expressed genes in the GSE38713 dataset, targets of ASI, and human TF and TF cofactors. (D) Six intersecting interactions were subjected to protein–protein interaction plotting (connections between two protein nodes represent evidence of interactions, and more connecting lines between the two indicate a closer relationship between the predicted interactions). (E) mRNA expression of WDR5 in NCM460 cells treated with ASI or not was examined using RT‐qPCR. (F) The protein expression of WDR5 in NCM460 cells treated with ASI was examined using western blot analysis. (G) Positive rate of WDR5 in the colonic tissues of mice detected by immunohistochemistry. The data are expressed as mean ± SEM, n = 3 (E, F) or 7 (G). ANOVA was applied for each comparison. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: After mixing with Protein A + G beads for 30 min at 4°C and centrifugation at 1000 g at 4°C, the supernatant was immunoprecipitated with antibodies against H3K4me3 (1:100, MBS8527387, MyBioSource) and WDR5 (1:50, 13,105, Cell Signaling Technologies, Beverly, MA, USA) at 4°C overnight.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Comparison

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Astragaloside IV Alleviates Ulcerative Colitis Progression by Inhibiting WDR5 ‐Mediated ENO1 H3K4me3 Modification

doi: 10.1002/kjm2.70064

Figure Lengend Snippet: Overexpression of WDR5 weakens the effects of ASI on DSS‐induced NCM460 cells. (A) Detection of mRNA expression of WDR5 in NCM460 cells treated with ASI + OE‐Vector/OE‐WDR5 was examined using RT‐qPCR. (B) The viability of NCM460 cells was examined using the CCK‐8 assay. (C) The apoptosis of NCM460 cells was examined using the TUNEL assay. (D) The protein expression of Cleaved‐Caspase‐3 in NCM460 cells was examined using Western blot assays. (E) The protein expression of ZO‐1 and claudin‐3 in NCM460 cells was examined using Western blot assays. (F) TNF‐α, IL‐6, IL‐1β, and IL‐10 contents in NCM460 cell culture supernatant were examined using ELISA. The data are expressed as the means ± SEM, n = 3. An unpaired t‐test (A) or ANOVA (B–F) was applied for each comparison. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: After mixing with Protein A + G beads for 30 min at 4°C and centrifugation at 1000 g at 4°C, the supernatant was immunoprecipitated with antibodies against H3K4me3 (1:100, MBS8527387, MyBioSource) and WDR5 (1:50, 13,105, Cell Signaling Technologies, Beverly, MA, USA) at 4°C overnight.

Techniques: Over Expression, Expressing, Plasmid Preparation, Quantitative RT-PCR, CCK-8 Assay, TUNEL Assay, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Comparison

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Astragaloside IV Alleviates Ulcerative Colitis Progression by Inhibiting WDR5 ‐Mediated ENO1 H3K4me3 Modification

doi: 10.1002/kjm2.70064

Figure Lengend Snippet: WDR5 promotes ENO1 transcription through H3K4me3 modification. (A) The intersection of differentially expressed genes in the GSE38713 dataset and WDR5 targets in the hTFtarget website. (B) The intersection of differentially expressed genes in the GSE38713 dataset, WDR5 targets in the hTFtarget website, and UC‐related genes downloaded from the GeneCards dataset. (C) ChIP‐seq analysis of H3K4me3 binding peaks on the ENO1 promoter region. (D) ChIP‐seq analysis of WDR5 binding peaks on the ENO1 promoter region. (E) Correlation analysis of WDR5 and ENO1 expression in the colon in the GEPIA dataset. (F) The protein expression of ENO1 in the colonic tissues of mice was detected by western blot analysis. (G) ENO1 mRNA expression in DSS‐stimulated NCM460 cells infected with a lentiviral vector overexpressing WDR5 was assessed using RT‐qPCR. (H) Enrichment of the ENO1 promoter region by anti‐WDR5 and anti‐H3K4me3 in NCM460 cells overexpressing WDR5 was analyzed using a ChIP assay. (I) H3K4me3 protein expression in DSS‐stimulated NCM460 cells infected with a lentiviral vector overexpressing WDR5 was examined using Western blot assay. The data are expressed as the means ± SEM, n = 3 (G–I) or 7 (F). An unpaired t ‐test (G–I) or ANOVA (F) was applied for each comparison. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: After mixing with Protein A + G beads for 30 min at 4°C and centrifugation at 1000 g at 4°C, the supernatant was immunoprecipitated with antibodies against H3K4me3 (1:100, MBS8527387, MyBioSource) and WDR5 (1:50, 13,105, Cell Signaling Technologies, Beverly, MA, USA) at 4°C overnight.

Techniques: Modification, ChIP-sequencing, Binding Assay, Expressing, Western Blot, Infection, Plasmid Preparation, Quantitative RT-PCR, Comparison

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Astragaloside IV Alleviates Ulcerative Colitis Progression by Inhibiting WDR5 ‐Mediated ENO1 H3K4me3 Modification

doi: 10.1002/kjm2.70064

Figure Lengend Snippet: Silencing of ENO1 alleviates DSS‐induced NCM460 cell injury in the presence of WDR5 overexpression. The mRNA and protein expression of ENO1 in NCM460 cells infected with OE‐WDR5 + sh‐NC/sh‐ENO1 was examined using RT‐qPCR (A) and western blot analysis (B). (C) The viability of NCM460 cells was examined using the CCK‐8 assay. (D) The apoptosis of NCM460 cells was examined using the TUNEL assay. (E) The protein expression of Cleaved‐Caspase‐3 in NCM460 cells was examined using Western blot assays. (F) TNF‐α, IL‐6, IL‐1β, and IL‐10 contents in NCM460 cells were examined using ELISA. The data are expressed as the means ± SEM, n = 3. An unpaired t ‐test was applied for each comparison. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: After mixing with Protein A + G beads for 30 min at 4°C and centrifugation at 1000 g at 4°C, the supernatant was immunoprecipitated with antibodies against H3K4me3 (1:100, MBS8527387, MyBioSource) and WDR5 (1:50, 13,105, Cell Signaling Technologies, Beverly, MA, USA) at 4°C overnight.

Techniques: Over Expression, Expressing, Infection, Quantitative RT-PCR, Western Blot, CCK-8 Assay, TUNEL Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Astragaloside IV Alleviates Ulcerative Colitis Progression by Inhibiting WDR5 ‐Mediated ENO1 H3K4me3 Modification

doi: 10.1002/kjm2.70064

Figure Lengend Snippet: Silencing of ENO1 mitigates the UC‐like symptom in mice in the presence of WDR5 overexpression. The mRNA and protein expression of ENO1 and WDR5 in colon tissues of mice treated with OE‐WDR5 + sh‐NC/sh‐ENO1 and ASI was examined using RT‐qPCR (A) and western blot analysis (B). (C) Positive rate of ENO1 and WDR5 in the colonic tissues of mice detected by immunohistochemistry. (D) Measurement of the length of the colon in mice at d 15 of modeling. (E) Analysis of colonic tissue pathology and tissue damage scores using HE staining. Detection of NO (F) and MPO content (G) in the colonic tissues of mice. The data are expressed as the means ± SEM, n = 7. ANOVA was applied for each comparison. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: After mixing with Protein A + G beads for 30 min at 4°C and centrifugation at 1000 g at 4°C, the supernatant was immunoprecipitated with antibodies against H3K4me3 (1:100, MBS8527387, MyBioSource) and WDR5 (1:50, 13,105, Cell Signaling Technologies, Beverly, MA, USA) at 4°C overnight.

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Staining, Comparison

Journal: The Kaohsiung Journal of Medical Sciences

Article Title: Astragaloside IV Alleviates Ulcerative Colitis Progression by Inhibiting WDR5 ‐Mediated ENO1 H3K4me3 Modification

doi: 10.1002/kjm2.70064

Figure Lengend Snippet: Possible mechanism of action of ASI in UC. Inhibition of WDR5‐mediated modification of H3K4me3 on the ENO1 promoter by ASI hinders transcriptional activation of ENO1 to alleviate symptoms of UC.

Article Snippet: After mixing with Protein A + G beads for 30 min at 4°C and centrifugation at 1000 g at 4°C, the supernatant was immunoprecipitated with antibodies against H3K4me3 (1:100, MBS8527387, MyBioSource) and WDR5 (1:50, 13,105, Cell Signaling Technologies, Beverly, MA, USA) at 4°C overnight.

Techniques: Inhibition, Modification, Activation Assay

Journal: Brain and Behavior

Article Title: Protective role of activating transcription factor 3 against neuronal damage in rats with cerebral ischemia

doi: 10.1002/brb3.2522

Figure Lengend Snippet: Activating transcription factor 3 (ATF3)/CCL2 regulates microglia activation in rats. (a) mRNA expression of pro‐inflammatory factors in rat brain tissues by reverse transcriptase quantitative polymerase chain reaction (RT‐qPCR). (b) mRNA expression of anti‐inflammatory factors in rat brain tissues by RT‐qPCR. (c) Immunofluorescence validation of ATF3 and CCL2 localization in microglia. (d) Immunohistochemical detection of IBA1 expression in rat brain tissues. (e) Assessment of microglia activation by immunofluorescence detection of BrdU/IBA1. Data are expressed as mean ± SD, and statistical significance was determined using one‐way analysis of variance (ANOVA) (a–e), followed by Tukey multiple comparison test. *#& p < .05

Article Snippet: The sections were blocked in 10% normal goat serum (ZLI‐9056; ZSGB‐Bio, Beijing, China) containing 0.3% Triton X‐100 for 60 min at room temperature and incubated with antibodies against ATF3 (1:150, ab191513; Abcam), neuronal nuclear antigen (NeuN, 1:200, ab104224; Abcam), IBA1 (1:200, NBP2‐19019; Novus Biologicals), and BrdU (1:300, ab8152, Abcam) overnight at 4°C.

Techniques: Activation Assay, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Immunofluorescence, Biomarker Discovery, Immunohistochemical staining, Comparison