Journal: Immunity, inflammation and disease

Article Title: Immune cell C/EBPβ deficiency is associated with hepatic mononuclear defects and spontaneous hepatitis but not steatohepatitis induced liver fibrosis.

doi: 10.1002/iid3.728

Figure Lengend Snippet: FIGURE 1 Altered hepatic myeloid cell populations in immune cell restricted C/EBPβ deficient mice. Single cell suspensions of hepatic cells were obtained 8 weeks after bone marrow transplantation of C/EBPβ−/−or C/EBPβ+/+ bone marrow into irradiated wild type recipients mice. Representative flow cytometry dot plots of singlets, CD45+ live (DAPI negative) cells including gating strategy for neutrophils, Ly6C+ monocytes, MHC II positive myeloid cells and Kupffer cells are depicted (A). Gating and representative dot plots of hepatic Ly6C−monocytes in indicated mice (B). Quantification of myeloid cell counts in the livers of indicated mice (C). Precent of TIM4+

Article Snippet: Anti‐mouse F4/80 (clone REA126) and anti‐mouse Tim4 (clone REA999) was purchased from Miltenyi Biotec.

Techniques: Transplantation Assay, Irradiation, Flow Cytometry

Journal: Journal of Cancer

Article Title: MUC15 is an independent prognostic factor that promotes metastases of MYCN non-amplified neuroblastoma.

doi: 10.7150/jca.89360

Figure Lengend Snippet: Figure 2. MUC15 is associated with poor prognosis in MYCN-NA NB from GEO datasets. A. The expression level of MUC15 across different INSS stages of NB (GSE45547 and GSE49710). B. Comparison of MUC15 expression between high-risk and non-high-risk NB cohorts (GSE49710). C. Differences in EFS and OS were evaluated

Article Snippet: The sections were then incubated overnight at 4°C with anti-MUC15 antibody (dilution of 1:100, Santa Cruz Biotechnology, USA).

Techniques: Expressing, Comparison

Journal: Journal of Cancer

Article Title: MUC15 is an independent prognostic factor that promotes metastases of MYCN non-amplified neuroblastoma.

doi: 10.7150/jca.89360

Figure Lengend Snippet: Figure 3. MUC15 is highly expressed in MYCN-NA NB. A. Expression of MUC15 in tissues with different stages. Blue and green fluorescence represent DAPI (nucleus) and MUC15, respectively. The histogram showed the relative fluorescence area (n=10). B. RT-PCR showed the MUC15 expression in patients with different stages (I+II, n=15; Ⅲ+Ⅳ, n=20). C. Western blotting revealed the ratio of MUC15 to internal reference protein in three different neuroblastoma cell lines. *p < 0.05, **p < 0.01.

Article Snippet: The sections were then incubated overnight at 4°C with anti-MUC15 antibody (dilution of 1:100, Santa Cruz Biotechnology, USA).

Techniques: Expressing, Fluorescence, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Journal of Cancer

Article Title: MUC15 is an independent prognostic factor that promotes metastases of MYCN non-amplified neuroblastoma.

doi: 10.7150/jca.89360

Figure Lengend Snippet: Figure 4. MUC15 promotes NB cell migration. A. Left: Transwell images of migration for NB cells randomly selected from each treatment group. Right: The bar chart showed the statistics of migration results of SK-N-AS and SK-N-SH. B. GESA analysis of migration-related pathways in the MYCN-NA subgroup of GSE45547. C. Western blotting showed increased expressions of three phosphorylated FAKs exposing to MUC15 overexpression and exogeneous MUC15 (20ng/ml) in SK-N-SH. D. Cytoskeletal protein staining in SK-N-AS. The green fluorescence for phalloidin combined with filamentous actin (F-actin) and excited with 488nm photoexcitation. The blue staining is DAPI for nuclear staining and MERGE as an overlapping image of two channels. SH#1&2 MUC15, knockdown of MUC15; OE-MUC15, overexpression of MUC15; MUC15, exogeneous MUC15 protein (20ng/ml). *p < 0.05, **p < 0.01, ****P<0.0001.

Article Snippet: The sections were then incubated overnight at 4°C with anti-MUC15 antibody (dilution of 1:100, Santa Cruz Biotechnology, USA).

Techniques: Migration, Western Blot, Over Expression, Staining, Fluorescence, Knockdown

Journal: Journal of Cancer

Article Title: MUC15 is an independent prognostic factor that promotes metastases of MYCN non-amplified neuroblastoma.

doi: 10.7150/jca.89360

Figure Lengend Snippet: Figure 5. MUC15 facilitates migration via MYCT1. A. Transcriptome analysis of MUC15 overexpression and control groups. B. MYCT1 expression situation after overexpression of MUC15 by RT-PCR in SK-N-SH (n=6). C. The content of MYCT1 after MUC15 overexpression in SK-N-SH was detected by western blotting. D. The correlation analysis between MUC15 and MYCT1 in GSE45547 and GSE49710. OE-MUC15, overexpression of MUC15; MUC15, exogeneous MUC15 protein (20ng/ml). *p < 0.05.

Article Snippet: The sections were then incubated overnight at 4°C with anti-MUC15 antibody (dilution of 1:100, Santa Cruz Biotechnology, USA).

Techniques: Migration, Over Expression, Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Journal of Cancer

Article Title: MUC15 is an independent prognostic factor that promotes metastases of MYCN non-amplified neuroblastoma.

doi: 10.7150/jca.89360

Figure Lengend Snippet: Figure 6. MYCT1 rescues the MUC15-induced migration of NB. A. Transwell images of migration for NB cells were randomly selected from each group treated as indicated. B. The statistics analysis of SK-N-AS and SK-N-SH in A. C. Western blotting showed the expressions of three FAK phosphorylation in SK-N-SH treated as indicated. OE-MUC15, overexpression of MUC15; OE-(MUC15+MYCT1), overexpression of MUC15 and MYCT1; MUC15, exogeneous MUC15 protein (20ng/ml); MUC15+OE-MYCT1, exogeneous MUC15 protein (20ng/ml) and overexpression of MYCT1; OE-MYCT1, overexpression of MYCT1. ****P<0.0001.

Article Snippet: The sections were then incubated overnight at 4°C with anti-MUC15 antibody (dilution of 1:100, Santa Cruz Biotechnology, USA).

Techniques: Migration, Western Blot, Phospho-proteomics, Over Expression

Journal: Frontiers in Medicine

Article Title: Urinary CD8+HLA-DR+ T Cell Abundance Non-invasively Predicts Kidney Transplant Rejection

doi: 10.3389/fmed.2022.928516

Figure Lengend Snippet: Gating strategies for T cell subsets (A) and tubular epithelial cells (TEC) (B) . Isotype controls are displayed as blue, while full stains are represented in red. (C.1) Schematic overview of investigated subsets. Proximal TECs were defined CD10+ and CD13+, while distal TECs were characterized being CD227+ and CD326(EpCAM)+. (C.2) Maturation of naïve T cells into memory T cells. (D) Workflow for epigenetic analysis of urine samples. SSC, side scatter; FSC, forward scatter; TNV, naïve T cells; TEM, T effector memory cells; TCM, T central memory cells; TEMRA, T effector memory cells re-expressing CD45RA.

Article Snippet: The following antibodies were used: for T cells anti-CD3-APCeF780 (eBioscience, SK7, mo IgG1k), -CD4-PEVio770 (Miltenyi Biotec, REA623, REA) -CD8-APC (Biolegend, SK1, mo IgG1k) -CD45RO-PE (Biolegend, UCHL1, mo IgG1k2), -CD45-BUV805 (BD, 3D12, rat IgG1ak), -CCR7-BV421 (Biolegend, G043H7, mo IgG2ak), -HLA-DR-BUV395 (BD, G46-6, mo IgG2ak), -CD28-FITC (Biolegend, CD28.2, mo IgG1k) and for tubular epithelial cells anti-Cytokeratin-FITC (Miltenyi Biotec, CK3-6H5, mo IgG1k), -Vimentin-APC (Miltenyi Biotec, REA409, REA), -CD10-PeVio770 (Miltenyi Biotec, REA877, REA), -CD13-APCVio770 (Miltenyi Biotec, REA263, REA), -CD227-PE (Miltenyi Biotec, REA448, REA), -CD326-BV711 (Biolegend, 9C4, mo IgG2b).

Techniques: Expressing

Journal: Nutrients

Article Title: Xenogeneic-Free Human Intestinal Organoids for Assessing Intestinal Nutrient Absorption

doi: 10.3390/nu14030438

Figure Lengend Snippet: The structures and properties of XF-HIOs. ( A ) A photograph of three-dimensional xenogeneic-free human intestinal organoids (XF-HIOs). ( B ) The level of expression of mRNAs for LGR5 , Villin , and CDX2 relative to that of the human small intestine. Data shown represent the mean ± S.E. values of three independent experiments. n.s.; not significant. ( C ) Immunofluorescent images of XF-HIOs with anti-VILLIN, CDX2, and MUCIN2 (MUC2). DAPI, 4′, 6-diamidino-2-phenylindole. Scale bars: 50 μm.

Article Snippet: Secreted Muc2 was assessed using Human Muc2 ELISA Kit (Elabscience, TX, USA).

Techniques: Expressing

Journal: Animals : an Open Access Journal from MDPI

Article Title: EGCG Derivatives Alleviate Diquat-Induced Liver and Gut Damage in Mice by Activating an Antioxidant Pathway and Enhancing Barrier Function

doi: 10.3390/ani16060966

Figure Lengend Snippet: Effects of EGCG derivatives on jejunal morphology and barrier function in mice. n = 6. ( A ) Jejunal villus height; ( B ) Jejunal crypt depth; ( C ) Villus height to crypt depth ratio; ( D – G ) The relative protein expression of ZO-1, Occludin, and MUC2 in the jejunum. T0, Control; T1, Diquat; T2, EGCG + Diquat; T3, Epigallocatechin octanoate + Diquat; T4, Epigallocatechin p -chloromethylbenzoate + Diquat; T5, Epigallocatechin ibuprofen ester + Diquat. a–d Different superscript letters indicate significant differences ( p < 0.05).

Article Snippet: The antibodies used were as follows: Nrf2 (#16396-1-AP), NQO1 (#11451-1-AP), MUC2 (#27675-1-AP) and Catalase (#21260-1-AP) from Proteintech (Wuhan, China); HO-1 (#AF5393), Occludin (#DF7504), and ZO-1 (#AF5154) from Affinity Biosciences Group Ltd. (Cincinnati, OH, USA). β-actin (#I102) served as the internal reference protein (Bioworld Biotech, Nanjing, China).

Techniques: Expressing, Control

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 overexpression on cellular and secreted mucin 5AC (MUC5AC) and mucin 5B (MUC5B) levels in NCI–H292 cells. A, NCI–H292 cells were transfected with integrin β1 (β1 cDNA) or control vectors (CNTL) and cultured on a 96-well plate. Integrin β1 and β-actin protein levels were detected in cells via immunoblotting with specific antibodies, and their levels were measured using ChemiDoc. B, MUC5AC levels in transfected cells were detected using the dot blot method and measured (cellular). C, MUC5AC levels were detected in the culture medium of transfected cells via the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± standard deviation [SD], n = 5). D, MUC5B levels in the transfected cells were detected using the dot blot method and measured (cellular). E, MUC5B levels in the culture medium of the transfected cells were detected using the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± SD, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular).

Techniques: Over Expression, Transfection, Control, Cell Culture, Western Blot, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin α2 overexpression on cellular and secreted mucin 5AC (MUC5AC) levels in NCI–H292 cells. A, NCI–H292 cells were transfected with integrin α2 (α2 cDNA) or control vectors (CNTL) and cultured on a 96-well plate. A, MUC5AC levels in transfected cells were detected using the dot blot method and measured (cellular). B, MUC5AC levels were detected in the culture medium of transfected cells via the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± standard deviation [SD], n = 5). Fold changes were based on control cells (mean ± SD, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular).

Techniques: Over Expression, Transfection, Control, Cell Culture, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 depletion on cellular and secreted mucin 5AC (MUC5AC) and ROS levels in NCI–H292 cells. A, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), and cultured on a 96-well plate. Integrin β1 and β-actin protein levels were detected via immunoblotting and measured using ChemiDoc. B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular). C, MUC5AC levels in the culture medium of the in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). D, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular). E, MUC5AC levels in the culture medium of the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). F, ROS levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected by hydrogen peroxide/peroxidase assay (cellular). Fold changes were based on control cells (mean ± standard deviation, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular).

Techniques: Transfection, Control, Cell Culture, Western Blot, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 depletion on cellular and secreted mucin 5B (MUC5B) levels in NCI–H292 cells. A, MUC5B levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL) transfected cells were detected via the dot blot method and measured (cellular). B, MUC5B levels in the culture medium of the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). C, MUC5B levels in the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (cellular). D, MUC5B levels in the culture medium of the integrin β1 siRNA (β1 siRNA, Thermo Fisher) or control siRNA (CNTL), transfected cells were detected via the dot blot method and measured (secreted). Fold changes were based on control cells (mean ± standard deviation, n = 5). Fold changes were normalized for cell numbers. Student's t-test was used to obtain p -values. Asterisks indicate statistical significance, * p < 0.05. Representative results for three independent experiments are presented.

Article Snippet: B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular).

Techniques: Control, Transfection, Dot Blot, Standard Deviation

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Effects of integrin β1 overexpression or depletion on cellular mucin 5AC (MUC5AC) and mucin 5B (MUC5B) levels in NCI–H292 cells. NCI–H292 cells (3.1 × 10 4 cells/cm 2 ) were transfected with integrin β1 vector (β1 cDNA), control vector (CNTL), integrin β1 siRNA (β1 siRNA), or control siRNA (CNTL), and cultured on Lab-Tek Chamber Slides for 48 h MUC5AC and MUC5B protein expression was detected via immunohistochemical analysis (green). Cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI, blue). Yellow bars indicate 100 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular).

Techniques: Over Expression, Transfection, Plasmid Preparation, Control, Cell Culture, Expressing, Immunohistochemical staining, Staining

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: Activities of extracellular signal-regulated kinase (ERK) in NCI–H292 cells transfected with integrin β1 cDNA (β1 cDNA) or siRNA (β1 siRNA). A, NCI–H292 cells (1 × 10 4 cells/well) were transfected with integrin β1 cDNA or siRNA, cultured, and sampled in a 96-well plate. Phosphorylated ERK and total ERK protein levels were detected. Representative results for three independent experiments are presented. B, A densitometry analysis of phosphorylated ERK/total ERK in NCI–H292 cells which were transfected with integrin β1 cDNA or siRNA. C, D, NCI–H292 cells were transfected with integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology), control siRNA (CNTL) and cultured with (+) or without (−) 10 μM of U0126 on a 96-well plate. The cellular (cellular, C) and secreted (secreted, D) MUC5AC levels were detected via the dot blot method and measured.

Article Snippet: B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular).

Techniques: Transfection, Cell Culture, Control, Dot Blot

Journal: Biochemistry and Biophysics Reports

Article Title: Integrin β1 subunit regulates cellular and secreted MUC5AC and MUC5B production in NCI–H292 human lung epithelial cells

doi: 10.1016/j.bbrep.2021.101124

Figure Lengend Snippet: A hypothetical model of integrin β1 reduces MUC5AC expression via ERK activation and ROS formation. The arrow-headed lines and bar-headed lines indicate activation and inhibition, respectively.

Article Snippet: B, MUC5AC levels in the integrin β1 siRNA (β1 siRNA, Santa Cruz Biotechnology) or control siRNA (CNTL), transfected cells were detected using the dot blot method and measured (cellular).

Techniques: Expressing, Activation Assay, Inhibition