rabbit anti icam1 antibody  (Proteintech)

Journal: bioRxiv

Article Title: Engineered human iPS cell models reveal altered podocytogenesis and glomerular capillary wall in CHD-associated SMAD2 mutations

doi: 10.1101/2024.08.02.606108

Figure Lengend Snippet: a , Schematic representation of the derivation of endothelial cells. b , Representative phase contrast images of the differentiated endothelial cells two additional culture days after MAC sorting. Scale bar, 275 µm. c , Endothelial cell differentiation yield calculated after MAC sorting. SMAD2 -/- and SMAD2 mh1/mh1 endothelial cells had a lower differentiation yield compared to the WT. Data are mean values ± SEM; n = 3 independent experiments. P -value was calculated by one-way ANOVA with post hoc Dunnett’s test. d , Representative immunofluorescent images showing the expression of endothelial marker Platelet endothelial cell adhesion molecule (PECAM1 /CD31, light grey) and Vascular Endothelial-Cadherin (VECAD , red). Single-channel images are shown next to the merged images for the entire field of view. DAPI, shown in blue, labels the nuclei, and the scale bar represent 100 µm. e , Representative immunofluorescent images showing the expression of endothelial marker PECAM1 (yellow) and mesenchymal marker Intercellular Adhesion Molecule 1 (ICAM1/CD54) in WT, SMAD2 -/- , and SMAD2 mh1/mh1 endothelial cells 2 weeks after differentiation. Single-channel images are shown next to the merged images for the entire field of view. DAPI, shown in light grey, labels the nuclei, and the scale bar represent 100 µm for all images. f , Representative immunoblots showing the expression of EC, EMT, and mesenchymal markers in the mutant endothelial cells, including VECAD, TWIST , and Transgelin . Data is representative of 2 biologically independent experiments.

Article Snippet: Primary antibodies used for Western blot were rabbit Anti-Phospho-SMAD1 (Ser206) (Cell Signaling; mAb #5753, 1:1000), rabbit Anti-SMAD2 (D43B4) (Cell Signaling; mAb #5339, 1:1000) rabbit Anti-SMAD3 (C67H9) (Cell Signaling; mAb #9523, 1:1000), rabbit Anti-Phospho-SMAD3 (C25A9) (Cell Signaling; mAb #9520, 1:1000), rabbit Anti-SMAD4 (D3R4N) (Cell Signaling; mAb #46535, 1:1000), rabbit Anti-SMAD6 (AbClonal; pAb #9520, 1:250), rabbit Anti-SMAD7 (AbClonal; pAb #16396, 1:250), rabbit Anti-Brachyury (Abcam; ab20680, 1:1000), rabbit Anti-CDX2 antibody (Abcam; ab76541, 1:1000), goat Anti-Goosecoid (R&D Systems, AF4086, 1:500), rabbit Anti-PAX2 (Cell Signaling; mAb #9666S, 1:1000), rabbit Anti β-Catenin (D10A8) (Cell Signaling; mAb #8480, 1:1000), mouse Anti-Twist antibody (Abcam; ab175430, 1:1000), rabbit Anti-Snail (C15D3) (Cell Signaling; mAb #3879, 1:1000), rabbit Anti-alpha smooth muscle Actin antibody (Abcam; mAb #ab7817, 1:500), rabbit Anti-MLKL antibody (Abcam; mAb #ab184718, 1:500), guinea pig Anti-Nephrin antibody (ARP; #GPN02, 1:500), rabbit Anti-Podocin (Abcam, mAb #ab50339, 1:1000), mouse Anti-Synaptopodin (D-9) (SantaCruz, mAb #sc-515842, 1:1000), mouse Anti-CD2AP (B-4) (SantaCruz, mAb #sc-25273, 1:1000), mouse Anti-GLEPP1 (B-6) (SantaCruz, mAb #sc-365354, 1:1000), mouse Anti-YAP (63.7) (SantaCruz; mAb #sc-101199, 1:1000), rabbit Anti-phospho-YAP (D9W2I) (Cell Signaling; mAb #13008, 1:1000), rabbit Anti-TEAD1 (D9X2L) (Cell Signaling; mAb #12292, 1:1000), rabbit Anti-CTGF (D8Z8U) (Cell Signaling; mAb #86641, 1:1000), rabbit Anti-Naked2 (C67C4) (Cell Signaling; mAb #2073, 1:1000), mouse Anti-VE-Cadherin (BV-9) (SantaCruz, mAb #sc-52751, 1:1000); rabbit Anti-transgelin (AbClonal, pAb, #A6760, 1: 1000), rabbit Anti-ICAM1/CD54 (AbClonal, pAb, #A5597, 1:1000), rabbit Anti-vimentin (AbClonal, pAb, #A2584, 1:1000), sheep Anti-PECAM1 (Bio-Rad, mAb, #CO.3E1D4, 1:1000), and rabbit Anti-GAPDH (Millipore Sigma; #ABS16, 1:10000).

Techniques: Cell Differentiation, Expressing, Marker, Western Blot, Mutagenesis

Journal: bioRxiv

Article Title: Engineered human iPS cell models reveal altered podocytogenesis and glomerular capillary wall in CHD-associated SMAD2 mutations

doi: 10.1101/2024.08.02.606108

Figure Lengend Snippet: a , RNA expression of mesenchymal markers ICAM1, Smoothelin, and Vimentin in WT and mutant endothelial cells. Data are expressed relative to WT after normalizing to GAPDH . Data are mean values ± SEM; n = 3 independent experiments. P -values were calculated by one-way ANOVA with post hoc Dunnett’s test. b , Representative immunofluorescent images showing the expression of mesenchymal markers Transgelin (cyan) and TWIST (red) in endothelial cells after 2 weeks of maintenance culture. Single-channel images are shown next to the merged images for the entire field of view. DAPI, shown in grey, labels the nuclei, and the scale bar represent 100 µm for all images.

Article Snippet: Primary antibodies used for Western blot were rabbit Anti-Phospho-SMAD1 (Ser206) (Cell Signaling; mAb #5753, 1:1000), rabbit Anti-SMAD2 (D43B4) (Cell Signaling; mAb #5339, 1:1000) rabbit Anti-SMAD3 (C67H9) (Cell Signaling; mAb #9523, 1:1000), rabbit Anti-Phospho-SMAD3 (C25A9) (Cell Signaling; mAb #9520, 1:1000), rabbit Anti-SMAD4 (D3R4N) (Cell Signaling; mAb #46535, 1:1000), rabbit Anti-SMAD6 (AbClonal; pAb #9520, 1:250), rabbit Anti-SMAD7 (AbClonal; pAb #16396, 1:250), rabbit Anti-Brachyury (Abcam; ab20680, 1:1000), rabbit Anti-CDX2 antibody (Abcam; ab76541, 1:1000), goat Anti-Goosecoid (R&D Systems, AF4086, 1:500), rabbit Anti-PAX2 (Cell Signaling; mAb #9666S, 1:1000), rabbit Anti β-Catenin (D10A8) (Cell Signaling; mAb #8480, 1:1000), mouse Anti-Twist antibody (Abcam; ab175430, 1:1000), rabbit Anti-Snail (C15D3) (Cell Signaling; mAb #3879, 1:1000), rabbit Anti-alpha smooth muscle Actin antibody (Abcam; mAb #ab7817, 1:500), rabbit Anti-MLKL antibody (Abcam; mAb #ab184718, 1:500), guinea pig Anti-Nephrin antibody (ARP; #GPN02, 1:500), rabbit Anti-Podocin (Abcam, mAb #ab50339, 1:1000), mouse Anti-Synaptopodin (D-9) (SantaCruz, mAb #sc-515842, 1:1000), mouse Anti-CD2AP (B-4) (SantaCruz, mAb #sc-25273, 1:1000), mouse Anti-GLEPP1 (B-6) (SantaCruz, mAb #sc-365354, 1:1000), mouse Anti-YAP (63.7) (SantaCruz; mAb #sc-101199, 1:1000), rabbit Anti-phospho-YAP (D9W2I) (Cell Signaling; mAb #13008, 1:1000), rabbit Anti-TEAD1 (D9X2L) (Cell Signaling; mAb #12292, 1:1000), rabbit Anti-CTGF (D8Z8U) (Cell Signaling; mAb #86641, 1:1000), rabbit Anti-Naked2 (C67C4) (Cell Signaling; mAb #2073, 1:1000), mouse Anti-VE-Cadherin (BV-9) (SantaCruz, mAb #sc-52751, 1:1000); rabbit Anti-transgelin (AbClonal, pAb, #A6760, 1: 1000), rabbit Anti-ICAM1/CD54 (AbClonal, pAb, #A5597, 1:1000), rabbit Anti-vimentin (AbClonal, pAb, #A2584, 1:1000), sheep Anti-PECAM1 (Bio-Rad, mAb, #CO.3E1D4, 1:1000), and rabbit Anti-GAPDH (Millipore Sigma; #ABS16, 1:10000).

Techniques: RNA Expression, Mutagenesis, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Mechanisms underlying neutrophils adhesion to triple-negative breast cancer cells via CD11b-ICAM1 in promoting breast cancer progression

doi: 10.1186/s12964-024-01716-5

Figure Lengend Snippet: Neutrophil CD11b directly binds to the tumor cell ICAM1, facilitating cell adhesion. A Flow analysis of CD11b in tumor infiltrating neutrophils from 4T1 or EMT6 tumor bearing mice at 2 weeks. B Flow analysis of ICAM1 in tumor cells from 4T1 or EMT6 tumor bearing mice at 2 weeks. C Quantitative analysis of ICAM1 expression in primary breast cancer of different molecular subtypes in TCGA database. D Pearson analysis of the correlations between ITGAM (CD11b) expression with ICAM1 expression in primary tumors with breast cancer from the TCGA database. E Pearson analysis of the correlations between ICAM1 expression with neutrophil infiltration in primary tumors with TNBC from the TCGA database. F Representative H&E and paired immunofluorescence staining images of neutrophils and ICAM1 in tumor from 4-week tumor bearing mice and TNBC patient. Red, ICAM1. Green, Ly6G (neutrophil in mice). MPO (neutrophil in human). Blue, DAPI. Scale bar, 500 μm and 100 μm respectively. G Principal component analysis (PCA) (left panel) and Volcano Plot (right panel) of RNA-seq of the tumor cell co-cultured with or without neutrophils. Each dot of Volcano Plot representing a gene and genes significantly upregulated in red and downregulated in green. H KEGG pathway analysis (left) and protein network analysis (right) of RNA-seq of differential gene in tumor cell co-cultured with or without neutrophils. Data are presented as the means ± SD from one representative experiment. Similar results were obtained from three independent experiments, unless indicated otherwise. Statistical analysis was performed by two-tailed unpaired Student's t test ( A , B ), Kruskal-wallis test ( C ). ns, not significant, * p ˂0.05, ** p ˂0.01, *** p ˂0.001, and **** p ˂0.0001

Article Snippet: Primary antibody, including Phospho-MAPK Family Antibody Sampler Kit (CST, Boston, MA, USA, #9910T), anti-P38 Rabbit mAb (CST, #8690T), SAPK/JNK Antibody (CST, #9252T), anti-ERK1+ERK2 antibody (Abcam, Cambridge, MA, USA, #ab184699), Anti-GAPDH (Huabio, Hangzhou, China, #SA30-01), anti-Beta Actin (Huabio, #B4-B2), HRP Conjugated Goat anti-Rabbit IgG Goat Polyclonal Antibody (Huabio, #HA1001), HRP Conjugated Goat anti-Mouse IgG Goat Polyclonal Antibody (Huabio, #HA1006), anti-ICAM1 Rabbit mAb (Abcam, #ab282575), anti-ICAM1 Rabbit mAb (Abcam, #ab222736) was applied.

Techniques: Expressing, Immunofluorescence, Staining, RNA Sequencing Assay, Cell Culture, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Mechanisms underlying neutrophils adhesion to triple-negative breast cancer cells via CD11b-ICAM1 in promoting breast cancer progression

doi: 10.1186/s12964-024-01716-5

Figure Lengend Snippet: Neutrophils enhance tumor invasion by acting on the tumor cell ICAM1 to promote the activation of the MAPK pathway in tumor cells. A Verification of the level of the ICAM1 protein in different tumor cell lines. shControl , tumor cells transfected with vector lentivirus. shICAM1-1 , shICAM1-2 , shICAM1-3 , tumor cells transfected with shICAM1 lentivirus of different target regions. B Levels of the MAPK pathway protein in different tumor cell lines co-cultured with or without neutrophils. C Analysis of the mRNA expression of MMP9 in different tumor cell lines co-cultured with or without neutrophils. D Invasion assay images and quantification of different tumor cell lines co-cultured with or without neutrophils. E Migration capability images and quantification of different tumor cell lines co-cultured with or without neutrophils. F Image (left panel) and tumor weights (right panel) of primary tumors from 4-week tumor bearing mice inoculated with shControl - or shICAM1 -4T1 cells. G Analysis of the mRNA expression of ICAM1 and MMP9 in primary tumors from 4-week tumor bearing mice inoculated with shControl - or shICAM1 -4T1 cells. H Levels of the MAPK pathway protein in primary tumors from 4-week tumor bearing mice inoculated with shControl - or shICAM1 -4T1 cells. I Representative H&E and paired immunofluorescence staining images of p-JNK and MMP9 in tumor from 4-week tumor bearing mice inoculated with shControl - or shICAM1 -4T1 cells. Red, p-JNK. Green, MMP9. Blue, DAPI. Scale bar, 100 μm. Data are presented as the means ± SD from one representative experiment. Similar results were obtained from three independent experiments, unless indicated otherwise. Statistical analysis was performed by two-tailed unpaired Student's t test (F, G) and one-way ANOVA (C, D, E). ns, not significant, * p ˂0.05, *** p ˂0.001, and **** p ˂0.0001

Article Snippet: Primary antibody, including Phospho-MAPK Family Antibody Sampler Kit (CST, Boston, MA, USA, #9910T), anti-P38 Rabbit mAb (CST, #8690T), SAPK/JNK Antibody (CST, #9252T), anti-ERK1+ERK2 antibody (Abcam, Cambridge, MA, USA, #ab184699), Anti-GAPDH (Huabio, Hangzhou, China, #SA30-01), anti-Beta Actin (Huabio, #B4-B2), HRP Conjugated Goat anti-Rabbit IgG Goat Polyclonal Antibody (Huabio, #HA1001), HRP Conjugated Goat anti-Mouse IgG Goat Polyclonal Antibody (Huabio, #HA1006), anti-ICAM1 Rabbit mAb (Abcam, #ab282575), anti-ICAM1 Rabbit mAb (Abcam, #ab222736) was applied.

Techniques: Activation Assay, Transfection, Plasmid Preparation, Cell Culture, Expressing, Invasion Assay, Migration, Immunofluorescence, Staining, Two Tailed Test

Journal: Cell Communication and Signaling : CCS

Article Title: Mechanisms underlying neutrophils adhesion to triple-negative breast cancer cells via CD11b-ICAM1 in promoting breast cancer progression

doi: 10.1186/s12964-024-01716-5

Figure Lengend Snippet: Atorvastatin can inhibit ICAM1 in tumor cells and reduce the malignant characteristics of TNBC. A Levels of the ICAM1 protein on tumor cell lines treated with atorvastatin for 24 hours in vitro . B Levels of the MAPK pathway protein in tumor cell lines co-cultured with or without neutrophils in the presence or absence of atorvastatin (4T1 10 μM, MDA-MB-231 5 μM) for 24 hours in vitro . C Analysis of the mRNA expression of MMP9 in tumor cell lines co-cultured with or without neutrophils in the presence or absence of atorvastatin for 24 hours in vitro . D Invasion assay images and quantification of tumor cell co-cultured with or without neutrophils in the presence or absence of atorvastatin for 24 hours in vitro . E Migration capability images and quantification of tumor cell lines co-cultured with or without neutrophils in the presence or absence of atorvastatin for 24 hours in vitro . F Image (left panel) and tumor weights (right panel) of primary tumors from 4-week tumor bearing mice inoculated with 4T1 cells after the intragastric administration of atorvastatin or PBS. G Analysis of the mRNA expression of ICAM1 and MMP9 in primary tumors from 4-week tumor bearing mice inoculated with 4T1 cells after the intragastric administration of atorvastatin or PBS. H Levels of the MAPK pathway protein in primary tumors from 4-week tumor bearing mice inoculated with 4T1 cells after the intragastric administration of atorvastatin or PBS. I Representative immunofluorescence staining images of neutrophils and ICAM1 in tumor from 4-week tumor bearing mice and triple negative patient treated with or without atorvastatin. Red, ICAM1. Green, Ly6G (neutrophil in mice). MPO (neutrophil in human). Blue, DAPI. Scale bar, 500 μm. J Representative immunofluorescence staining images of p-JNK and MMP9 in tumor from 4-week tumor bearing mice and triple negative patient treated with or without atorvastatin. Red, p-JNK. Green, MMP9. Blue, DAPI. Scale bar, 100 μm. Data are presented as the means ± SD from one representative experiment. Similar results were obtained from three independent experiments, unless indicated otherwise. Statistical analysis was performed by two-tailed unpaired Student's t test (F, G, J) and one-way ANOVA (C, D, E). ns, not significant, * p ˂0.05, ** p ˂0.01, *** p ˂0.001, and **** p ˂0.0001

Article Snippet: Primary antibody, including Phospho-MAPK Family Antibody Sampler Kit (CST, Boston, MA, USA, #9910T), anti-P38 Rabbit mAb (CST, #8690T), SAPK/JNK Antibody (CST, #9252T), anti-ERK1+ERK2 antibody (Abcam, Cambridge, MA, USA, #ab184699), Anti-GAPDH (Huabio, Hangzhou, China, #SA30-01), anti-Beta Actin (Huabio, #B4-B2), HRP Conjugated Goat anti-Rabbit IgG Goat Polyclonal Antibody (Huabio, #HA1001), HRP Conjugated Goat anti-Mouse IgG Goat Polyclonal Antibody (Huabio, #HA1006), anti-ICAM1 Rabbit mAb (Abcam, #ab282575), anti-ICAM1 Rabbit mAb (Abcam, #ab222736) was applied.

Techniques: In Vitro, Cell Culture, Expressing, Invasion Assay, Migration, Immunofluorescence, Staining, Two Tailed Test