Fig 1: VEGF blockade in combination with anti–CTLA-4 therapy increases T cell infiltration and polarizes macrophages to an immunostimulatory phenotype.FFPE 4T1 tumors were assessed for CD31 and NG2 (A); CD31 and ICAM-1 (B); T cell markers CD3 (C), CD8 (D), and FoxP3 (E); as well as macrophage markers F4/80 (F), iNOS (G), and Arg-1 (H). Slides were scanned and images were analyzed using NIS Elements (Nikon) and Fiji software. Representative images are shown with CD31 in red and other markers in brown. Scale bar, 50 μm. Quantification is shown to the right. Data are displayed as mean ± SEM (n = 4–6/group). *, P < 0.05; **, P < 0.005, by ANOVA with Tukey’s MCT.
Fig 2: Apremilast suppressed the expression of several adhesive molecules, chemokines, and chemokine receptors in chronic UC. (A) The mRNA expression of multiple chemokine receptors in inflamed colonic tissues. (B) Representative immunofluorescent staining of CCR5 (200× magnification) in colon sections. Scale bar: 100 µm. (C) The mRNA expression of several chemokines in inflamed colonic tissues. (D) The mRNA expression of Icam-1, Madcam-1, and Cd62e in inflamed colonic tissues. (E) Representative immunofluorescent staining of ICAM-1 (200× magnification) in colon sections. Scale bar: 100 µm. (F) Western blot assay of adhesive molecules and chemokine receptors in inflamed colonic tissues. (G) Western blot assay of ICAM-1 and CD62E expression in HT-29 cells. (H) IP-10 and IL-8 levels in TNF-a-cultured HT-29 cells, detected by ELISA. (I) The counts of Jurkat T, U937, and THP-1 cells chemotactic to the lower chambers. (A)–(F) Data are presented as mean ± SEM; n = 10 mice per group. (G) Data are presented of three independent experiments. (H, I) Data are presented as mean ± SEM (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 3: High expression of lncRNA HIF1A-AS2 and ATF2 are identified in atherosclerotic mice. (A) Body weight and composition of ApoE-/- mice fed with ND or HFD. (B) Measurement of the food intake of ApoE-/- mice fed with ND or HFD. (C) Lipid deposition in the aorta observed after Oil red O staining. (D) The thoracic aortic injury area detected by HE staining (100×). (E) Lipid deposition in the thoracic aorta observed after Oil red staining (100×). (F) ELISA detection of TNF-a, IL-1ß, and IL-6 levels in the serum of the ApoE-/- mice fed with ND or HFD. (G) Immunohistochemical detection of VCAM-1, ICAM-1, and MCP-1 expression in arterial tissues of the ApoE-/- mice fed with ND or HFD (400×). (H) The expressions of lncRNA HIF1A-AS2 and ATF2 in arterial tissues of the ApoE-/- mice fed with ND or HFD determined by RT-qPCR. (I) Correlation analysis of lncRNA HIF1A-AS2 expression and ATF2 expression. (J) The expression of ATF2 in arterial tissues of the ApoE-/- mice fed with ND or HFD detected by immunohistochemistry (400×). *p < 0.05 vs. the ApoE-/- mice fed with ND. Data (mean ± standard deviation) between two groups were analyzed using unpaired t test. n = 20 for mice fed with ND or HFD.
Fig 4: Aleglitazar reduces stroke damage and expression of inflammatory molecules after mild transient brain ischemia. Treatment with aleglitazar was begun on the day on which brain ischemia was induced. Daily treatment with aleglitazar was continued until sacrifice (a, b). Infarct size at 7 days after 30 min MCAo/reperfusion was assessed using NeuN-stained coronal brain sections. N = 6–8 mice per group. One-way ANOVA followed by Tukey’s multiple comparison test. *p < 0.05 versus vehicle-treated control mice. c Aleglitazar improves sensorimotor outcome after 30 min MCAo/reperfusion. N = 3–7 mice per group. Two-way ANOVA followed by Tukey’s multiple comparison test within each time point. *p < 0.05 and **p < 0.01 aleglitazar versus vehicle, ##p < 0.01 and ###p < 0.001 MCAo versus sham. d Aleglitazar provides neuroprotection against oxygen–glucose deprivation (OGD). Primary cortical neurons were pretreated with aleglitazar or vehicle for 24 h. Neuronal injury was assessed by measuring lactate dehydrogenase (LDH) release into the cell culture medium 24 h after OGD. MK-801 served as a positive control. N = 5 independent cultures per condition. Two-way ANOVA followed by Tukey’s multiple comparison test. ****p < 0.0001 relative to sham cultures; #p < 0.05, ##p < 0.01 for the effect of treatment with aleglitazar within the OGD condition. e Expression of inflammatory and angiogenesis-related genes in the ipsilateral, i.e., ischemic MCA territory at 7 days. Relative mRNA expression is reported as the value normalized to receptor accessory protein 5 (Reep5). N = 4–5 mice per group. Two-way ANOVA followed by Tukey’s multiple comparison test. *p < 0.05 relative to sham. #p < 0.05 relative to vehicle-treated MCAo animals. f IL-1ß and KC/GRO-alpha (CXCL1) concentrations were also measured in serum at 7 days. N = 4–5 mice per group. Two-way ANOVA followed by Tukey’s multiple comparison test. #p < 0.05 relative to vehicle-treated MCAo animals. g VCAM-1 and ICAM-1 Western blots were performed at 7 days. N = 3 mice per group. Two-way ANOVA followed by Tukey’s multiple comparison test. *p < 0.05 and **p < 0.01 relative to sham. #p < 0.05 and ##p < 0.01 relative to vehicle-treated MCAo animals
Fig 5: Selective hACE2 expression in OE and neurons is sufficient to confer hypoxemia and death following SARS-CoV-2 infection.(A-C) Immunohistochemistry of WT and Foxg1Cre/+;LSL-hACE2+/0 mouse OE (A), OB, and cerebral cortex (B) and lung (C) 6 days after infection with 105 PFU of SARS-CoV-2 using antibodies that recognize viral nucleocapsid, sustentacular cell E-cadherin, olfactory sensory neuron βIII-tubulin, neuronal NeuN, glial GFAP, AT1 PDPN, or AT2 DC-LAMP. Asterisk indicates OB with nucleocapsid staining. Arrows indicate nucleocapsid staining colocalized with βIII-tubulin+ OSNs (OE in A) or NeuN+ neurons (OB and cerebral cortex in B). White dotted lines trace the border between the OE and OB. Representative of n = 5 per genotype. (D) HE staining of WT and Foxg1Cre/+; hACE2fl/y lung tissue 6 days after exposure to 105 PFU of SARS-CoV-2 virus. Arrows, sites of focal consolidation. Asterisks, intravascular thrombi. Representative of n = 5 animals per genotype. (E, F) Immunohistochemistry of WT and Foxg1Cre/+; hACE2fl/y lung tissue 6 days after exposure to 105 PFU of SARS-CoV-2 virus using antibodies against ICAM-1 and PDPN, or vWF and PECAM. Arrows in I identify vWF-positive microvasculature of the lung in Foxg1Cre/+; hACE2fl/y animals. Representative of n = 5 animals per genotype. (G, H) Weight loss and survival of LSL-hACE2+/0 and Foxg1Cre/+; LSL-hACE2+/0 mice after infection with 105 or 104 PFU of SARS-CoV-2. N = 4 (LSL-hACE2+/0), 8 (Foxg1Cre/+; LSL-hACE2+/0 105 PFU), 5 (Foxg1Cre/+; LSL-hACE2+/0 104 PFU) and two independent experiments. Asterisks indicate time points at which significant differences in weight (C) or survival (D) were observed between infected Foxg1Cre/+; LSL-hACE2+/0 and LSL-hACE2+/0 mice animals. (I) Pulse oximetry of WT (LSL-hACE2+/0 or Foxg1Cre/+) and Foxg1Cre/+; LSL-hACE2+/0 mice infected with 104 viral titer at the time of harvest (Day 6–8). ***p < 0.001; ****p < 0.0001 determined by unpaired, two-tailed t test or log-rank Mantel Cox test. Scale bars in all images 50 μm. Numerical data in corresponding S1 Metadata tab. hACE2, human ACE2; ICAM-1, intracellular adhesion marker 1; OB, olfactory bulb; OE, olfactory epithelium; OSN, olfactory sensory neuron; PDPN, Podoplanin; PECAM, platelet endothelial cell adhesion molecule; PFU, plaque-forming unit; SARS-CoV-2, Severe Acute Respiratory Syndrome Coronavirus 2; vWF, von Willebrand’s Factor; WT, wild-type.
Supplier Page from Abcam for Anti-ICAM1 antibody [EPR16608]