Journal: Scientific Reports

Article Title: Loss of CEACAM1, a Tumor-Associated Factor, Attenuates Post-infarction Cardiac Remodeling by Inhibiting Apoptosis

doi: 10.1038/srep21972

Figure Lengend Snippet: ( A ), Raw data of serum CEACAM1 in patients with AMI (n = 26) and healthy controls (n = 24). ( B ), Logarithmically-transformed serum CEACAM1 data for patients with AMI.

Article Snippet: To detect expression of CEACAM1 and cleaved caspase-3 in the myocardium, sections of the middle LV slice were incubated overnight at 4 °C with rabbit anti-CEACAM1 antibody (Santa Cruz Biotechnology, Dallas, USA) or rabbit anti-cleaved caspase-3 antibody (Abcam, Cambridge, UK).

Techniques: Transformation Assay

Journal: Scientific Reports

Article Title: Loss of CEACAM1, a Tumor-Associated Factor, Attenuates Post-infarction Cardiac Remodeling by Inhibiting Apoptosis

doi: 10.1038/srep21972

Figure Lengend Snippet: ( A ), Time course of CEACAM1 mRNA level in sham, non-infarct area and infarct area. * P < 0.05 vs. sham 6h. Expression of CEACAM1 mRNA ( B ) and protein ( C ) in the left ventricle of wild-type (WT) mice with MI for 3 days (n = 4 in each group). * P < 0.01 vs. sham. ( D ), Immunohistochemical staining shows elevated myocardial CEACAM1 expression in mice with MI than in mice with sham operation (non-infarct area). ( E ), Immunoblotting of CEACAM1 expression in neonatal rat cardiomyocytes exposed to hypoxia for 24 hours (n = 5 in each group). * P < 0.01 vs. normoxia. ( F ), The CEACAM1 mRNA level in neonatal rat cardiomyocytes exposed to hypoxia (for 3h), angiotensin II (AngII, 1 umol/L), endothelin 1(0.1 umol/L) and H 2 O 2 (0.15 umol/L) for 24 hours. * P < 0.05 vs. normoxia.

Article Snippet: To detect expression of CEACAM1 and cleaved caspase-3 in the myocardium, sections of the middle LV slice were incubated overnight at 4 °C with rabbit anti-CEACAM1 antibody (Santa Cruz Biotechnology, Dallas, USA) or rabbit anti-cleaved caspase-3 antibody (Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemical staining, Staining, Western Blot

Journal: Scientific Reports

Article Title: Loss of CEACAM1, a Tumor-Associated Factor, Attenuates Post-infarction Cardiac Remodeling by Inhibiting Apoptosis

doi: 10.1038/srep21972

Figure Lengend Snippet: ( A ), Wild-type (WT) and CEACAM1 knockout (KO) mice were subjected to sham operation or MI. Then survival was monitored for 8 weeks. ( B ) Masson trichromatic staining of WT and KO mouse hearts at 8 weeks after MI (blue indicates collagen). ( C ) Quantification of the length of fibrosis in the infarct area (relative to the total LV circumference) and the fibrotic area in the border zone and the remote area. * P < 0.05 vs. the corresponding WT group, n = 6 in each group. ( D ) Representative echocardiographic images from the four groups. ( E ) Quantification of left ventricular end-systolic and end-diastolic diameter (LVESd, LVEDd), and left ventricular fractional shortening (LVFS). * P < 0.01 vs. corresponding sham group, # P < 0.05 vs. WT MI. (n = 4 in sham groups; n = 9 in MI groups).

Article Snippet: To detect expression of CEACAM1 and cleaved caspase-3 in the myocardium, sections of the middle LV slice were incubated overnight at 4 °C with rabbit anti-CEACAM1 antibody (Santa Cruz Biotechnology, Dallas, USA) or rabbit anti-cleaved caspase-3 antibody (Abcam, Cambridge, UK).

Techniques: Knock-Out, Staining

Journal: Scientific Reports

Article Title: Loss of CEACAM1, a Tumor-Associated Factor, Attenuates Post-infarction Cardiac Remodeling by Inhibiting Apoptosis

doi: 10.1038/srep21972

Figure Lengend Snippet: ( A ). TUNEL staining of cardiomyocytes exposed to normoxia or hypoxia in the presence/absence of recombinant human CEACAM1(rhCEACAM1). Scale bar, 50 μm. ( B ), Quantitation of TUNEL-positive cells in the four groups shown in A. * P < 0.01 vs. normoxia, and # P < 0.01 vs. hypoxia, n = 6 in each group. ( C ), Representative pictures of TUNEL-stained cultured cardiomyocytes exposed to normoxia or hypoxia in the presence/absence of lentivirus carrying si-RNA for CEACAM1. Scale bar, 20 μm.( D ), Quantitation of TUNEL-positive cells in the 6 groups shown in C. * P < 0.01 vs. normoxia, # P < 0.05 vs. hypoxia +negative control-siRNA (si-NC), n = 10 in each group.

Article Snippet: To detect expression of CEACAM1 and cleaved caspase-3 in the myocardium, sections of the middle LV slice were incubated overnight at 4 °C with rabbit anti-CEACAM1 antibody (Santa Cruz Biotechnology, Dallas, USA) or rabbit anti-cleaved caspase-3 antibody (Abcam, Cambridge, UK).

Techniques: TUNEL Assay, Staining, Recombinant, Quantitation Assay, Cell Culture, Negative Control

Journal: Scientific Reports

Article Title: Loss of CEACAM1, a Tumor-Associated Factor, Attenuates Post-infarction Cardiac Remodeling by Inhibiting Apoptosis

doi: 10.1038/srep21972

Figure Lengend Snippet: Cleaved caspase-3, mitochondrial Bax (Mito Bax), and cytosolic cytochrome C (Cyto C) were analyzed by Western blotting under either normoxic or hypoxic conditions. CoxIV and β-actin served as internal controls. ( A ) Representative western blots of the target and loading control proteins from cardiomyocytes with/without recombinant human CEACAM1 (rhCEACAM1). ( B ) Quantitative analysis of the protein expression shown in A. * P < 0.05 vs. normoxia, # P < 0.05 vs. hypoxia, n = 5 in each group. ( C ) Western blotting of cleaved caspase-3, mitochondrial Bax, and cytosolic cytochrome C in cardiomyocytes treated with negative control siRNA (si-NC) or siRNA for CEACAM1 (si-CEACAM1). * P < 0.01 vs. normoxia, # P < 0.05 vs. hypoxia+ si-NC, n = 5 in each group. ( E ) Quantitative analysis of GRP78 and CHOP mRNA levels from cardiomyocytes with/without recombinant human CEACAM1 (rhCEACAM1). * P < 0.05 vs. normoxia, # P < 0.05 vs. hypoxia, n = 5 in each group. ( F ) The GRP78 and CHOP mRNA levels in cardiomyocytes treated with si-NC or si-CEACAM1. * P < 0.01 vs. normoxia, # P < 0.05 vs. hypoxia+ si-NC, n = 5 in each group.

Article Snippet: To detect expression of CEACAM1 and cleaved caspase-3 in the myocardium, sections of the middle LV slice were incubated overnight at 4 °C with rabbit anti-CEACAM1 antibody (Santa Cruz Biotechnology, Dallas, USA) or rabbit anti-cleaved caspase-3 antibody (Abcam, Cambridge, UK).

Techniques: Western Blot, Control, Recombinant, Expressing, Negative Control

Journal: Scientific Reports

Article Title: Loss of CEACAM1, a Tumor-Associated Factor, Attenuates Post-infarction Cardiac Remodeling by Inhibiting Apoptosis

doi: 10.1038/srep21972

Figure Lengend Snippet: Both in vitro and in vivo , CEACAM1 induces mitochondrial translocation of Bax and mitochondria dysfunction with consequent activation of the cytochrome C-caspase-3 apoptotic signaling pathway, which promotes hypoxia-induced apoptosis and of post-infarction cardiac remodeling. In addition, CEACAM1 promotes hypoxia-induced cardiomyocyte apoptosis through GRP78 and CHOP pathway.

Article Snippet: To detect expression of CEACAM1 and cleaved caspase-3 in the myocardium, sections of the middle LV slice were incubated overnight at 4 °C with rabbit anti-CEACAM1 antibody (Santa Cruz Biotechnology, Dallas, USA) or rabbit anti-cleaved caspase-3 antibody (Abcam, Cambridge, UK).

Techniques: In Vitro, In Vivo, Translocation Assay, Activation Assay

Journal: Nature Communications

Article Title: An immune checkpoint score system for prognostic evaluation and adjuvant chemotherapy selection in gastric cancer

doi: 10.1038/s41467-020-20260-7

Figure Lengend Snippet: Multi-colour immunofluorescence staining of anti-CKpan and CD45 antibodies with CD155, NECTIN2, CEACAM1, HMGB1, SIGLEC6 or CD44 antibodies. Scale bar = 200 μm.

Article Snippet: Based on the collection and review of relevant literature (Supplementary Table ), we selected 20 immune checkpoints for immunohistochemical staining analysis: CD73 (ab175396, Abcam, 1 : 200), Galectin-9 (54330 S, Cell Signaling Technology (CST), 1 : 800), HMGB1 (ab18256, Abcam, 1 : 1000), FAS-L (ab186671, Abcam, 1 : 200), SIGLEC6 (ab38581, Abcam, 1 : 200), SIGLEC15 (ab174723, Abcam, 1 : 200), TLR4 (ab13556, Abcam, 1 : 200), ADENOSINE (ab40002, Abcam, 1 : 250), CEACAM1 (44464S, CST, 1 : 400), NECTIN2 (95333S, CST, 1 : 200), CD44 (3570S, CST, 1 : 50), CD155 (81254 S, CST, 1 : 200), VISTA (54979S, CST.

Techniques: Immunofluorescence, Staining

Journal: The Journal of Clinical Investigation

Article Title: Functional monocytic myeloid-derived suppressor cells increase in blood but not airways and predict COVID-19 severity

doi: 10.1172/JCI144734

Figure Lengend Snippet: (A) Gating strategy to identify M-MDSCs by flow cytometry. From live, single CD45+ leukocytes, cells expressing lineage markers (CD3, CD19, CD20, CD56, CD66abce) and HLA-DR were excluded and CD14+ M-MDSCs identified. (B) M-MDSC frequency per live CD45+ cells in blood and NPAs. HCs (blue): n = 12 (blood), n = 7 (NPAs). Patients with influenza (open circles): n = 19 (blood), n = 9 (NPAs). COVID-19 patients (solid circles): n = 140 (blood), n = 28 (NPAs). The dots are color-coded according to peak disease severity. (C) Peak frequency of blood M-MDSCs per live CD45+ cells across disease severity. HCs (blue): n = 12. Patients with COVID-19 (color-coded by peak disease severity): mild, n = 19; moderate, n = 53; severe, n = 56; fatal, n = 12. (D) Blood M-MDSC frequencies over time in patients with COVID-19: mild, n = 17; moderate, n = 53; severe, n = 56; fatal, n = 12. Line shows the locally estimated scatterplot smoothing (LOESS) with shaded 95% CI (fatal group wide CI, not presented). (E) Frequency of blood M-MDSCs in paired acute and convalescent samples from patients with COVID-19 (n = 6). (F) M-MDSC frequency in blood, NPA, and ETA samples from patients with severe (red, n = 16) and fatal (gray, n = 4) COVID-19. (G–I) Surface expression of (G) CD62L, (H) CD86, and (I) CCR2 on M-MDSCs in blood, NPAs, and ETAs from HCs (blue, NPAs n = 7, PBMCs n = 11) and COVID-19 patients (black, NPAs n = 25, ETAs n = 19, PBMCs n = 69). (J) Frequency of PMN-MDSCs of live CD45+ cells in blood from patients with COVID-19. HCs: n = 12. Patients with COVID-19: mild, n = 11; moderate, n = 47; severe, n = 42; and fatal, n = 8. (K) Frequency of blood PMN-MDSCs in paired acute and convalescent samples from patients with COVID-19 (n = 6). (B, C, and F–J) Comparisons of M-MDSC frequencies were performed using the nonparametric Kruskal-Wallis test with Dunn’s post hoc multiple-comparison test. In the strip charts, group medians are presented as horizontal lines and individual patients as jitter points.

Article Snippet: If a sufficient number of cells were available, a second staining was performed using antibodies against CD3 (SP34-2; BD), CD4 (L200; BD), CD11c (B-ly6; BD), CD14 (M5E2; BD), CD16 (3G8; BD), CD19 (SJ25-C1; Thermo Fisher Scientific), CD45 (HI30; BD), CD56 (HCD56; BioLegend), CD66abce (TET2; Miltenyi Biotec), CD123 (7G3; BD), LOX-1 (15C4; BioLegend), and HLA-DR (L243; BioLegend).

Techniques: Flow Cytometry, Expressing, Comparison, Stripping Membranes