Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Transcriptomic features of ex vivo CD4 + T RM cells from CD patients. ( A ) Representative flow cytometry of CD4 + CD45RO + CD69 + CD103 + lamina propria T cells in control and CD patients; percentages in right upper quadrant indicate CD69 + CD103 + T RM cells frequency. ( B ) Statistical analysis results for frequency of CD69 + CD103 + T RM cells among CD4 + CD45RO + lamina propria T cells. ( C ) Immunofluorescence staining of CD103 ( green ) and CD69 ( red ) in intestinal lamina propria tissues from control and CD patients; white arrows indicate double-positive cells. Scale bars represent 50 μm (left panel) and 25 μm (right zoom-in panel). ( D ) Representative flow cytometry of CD69 + CD103 + cells among isolated CD4 + T RM cells in control and CD patients; percentages in right upper quadrant indicate CD69 + CD103 + cell frequency. ( E ) Statistical analysis results for frequency of CD69 + CD103 + cells among isolated CD4 + T RM cells from control and CD patients. ( F ) PCA of RNA sequencing data from ex vivo CD4 + T RM cells in control and CD patients. ( G ) Volcano plot of differentially expressed genes in ex vivo CD4 + T RM cells from control and CD patients ( P < .05, ∣Log 2 (fold change)∣> 1). All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with unpaired two-sided t test in ( B ). ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Ex Vivo, Flow Cytometry, Control, Immunofluorescence, Staining, Isolation, RNA Sequencing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Transcriptomic features of ex vivo CD4 + T RM cells from CD patients. ( A ) Heatmap of top 40 genes that were up-regulated in CD4 + T RM cells from CD patients; color scale demonstrates Log 2 (fold change of gene expression level in CD versus control). ( B and C ) Top 10 up-regulated Gene Ontology and KEGG pathway clusters of differentially expressed genes in CD4 + T RM cells from CD and control patients; enriched pathways are shown as gene number and - Log 10 (false discovery rate q value). ( D ) Representative GSEA results of differentially expressed genes in CD4 + T RM cells from CD and control patients. ( E and F ) Representative GSEA results of differentially expressed genes in CD4 + T RM cells from CD and control patients. All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with unpaired two-sided t test in ( B ). ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Ex Vivo, Gene Expression, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Metabolomic analysis of ex vivo CD4 + T RM Cells in CD patients. Heatmap of metabolomic data from ex vivo CD4 + T RM cells in CD and control patients (n = 8).

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Ex Vivo, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Metabolomic analysis of ex vivo CD4 + T RM cells in CD patients. ( A and B ) The 2-dimensional and 3-dimensional PCA results of metabolomic data from ex vivo CD4 + T RM cells in CD and control patients (n = 8). ( C and D ) Enrichment analysis and KEGG pathway analysis of metabolites in CD4 + T RM cells from CD and control patients. ( E ) Volcano plot of metabolites in ex vivo CD4 + T RM cells from CD and control patients ( P < .05, ∣fold change∣> 1.5). ( F ) Heatmap of metabolites in fatty acid degradation pathway from CD4 + T RM cells of CD and control patients.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Ex Vivo, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Metabolite and gene expression level in fatty acid uptake and oxidation pathway from CD4 + T RM cells. ( A ) Labeling pattern of acetyl-CoA, aspartate, and TCA cycle intermediates (M+2) in the isotope tracing ( 13 C 16 -palmitic acid) experiment. Schematic model shows 13 C 16 -palmitic acid experiment in ex vivo CD4 + T RM cells from CD and control patients. Red circles depict carbons from 13 C 16 -palmitic acid, and black circles indicate unlabeled carbons. ( B ) Uptake of 13 C 16 -palmitic acid in ex vivo CD4 + T RM cells from CD and control patients. ( C ) FPKM value of CD36 and FABP5 from RNA sequencing data in ex vivo CD4 + T RM cells. ( D and E ) Protein level of CD36 and FABP5 in ex vivo CD4 + T RM cells from CD and control patients. ( F ) FPKM value of key enzymes involved in fatty acid biosynthesis from RNA sequencing data in ex vivo CD4 + T RM cells. All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with unpaired two-sided t test in A , B , C , E , and F . ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Gene Expression, Labeling, Ex Vivo, Control, RNA Sequencing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Lipidomic analysis of ex vivo CD4 + T RM cells in CD patients. ( A ) Heatmap of lipidomic data from ex vivo CD4 + T RM cells in CD and control patients (n = 4). ( B and C ) The 2-dimensional and 3-dimensional PCA plots of lipidomic data from ex vivo CD4 + T RM cells in CD and control patients (n = 4).

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Ex Vivo, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Lipidomic analysis of ex vivo CD4 + T RM cells in CD patients. ( A and B ) Bar chart depicts relative abundance of DAG and TAG levels in ex vivo CD4 + T RM cells from control and CD patients (n = 4). ( C ) Representative images of lipid droplets by Bodipy staining ( green ) in CD4 + T RM cells from CD and control patients. Scale bar, 25 μm. ( D and E ) Representative flow cytometry analysis of lipid droplets by Bodipy staining in CD4 + T RM cells from CD and control patients. ( F ) FPKM value of key enzymes involved in lipid lipolysis and lipophagy from RNA sequencing data in ex vivo CD4 + T RM cells. ( G ) The qPCR results of genes from lipid lipolysis and lipophagy pathway in CD4 + T RM cells. All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with unpaired two-sided t test in A , B , E , F , and G . ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Ex Vivo, Control, Staining, Flow Cytometry, RNA Sequencing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Transcription factor NF-κB regulates expression levels of lipid metabolism genes in CD4 + T RM cells. ( A and B ) Heatmap of genes in FAO pathway and NF-κB signaling pathway from RNA sequencing data in ex vivo CD4 + T RM cells of CD and control patients. ( C and D ) The qPCR results of genes in lipid metabolism and NF-κB signaling pathway from CD4 + T RM cells. All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with unpaired two-sided t test in C and D . ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Expressing, RNA Sequencing, Ex Vivo, Control

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Transcription factor NF-κB regulates expression levels of lipid metabolism genes in CD4 + T RM cells. ( A ) Western blot for nuclear and cytoplasmic p65 expression in CD4 + T RM cells from CD and control patients, tubulin, and lamin A were used as cytoplasmic and nuclear internal control. ( B ) Statistical analysis was performed with Mann-Whitney test. ( C ) Immunofluorescence staining of nuclear and cytoplasmic p65 in CD4 + T RM cells from CD and control patients (p65, green ; DAPI, blue ). Scale bar, 25 μm. ( D ) The mRNA levels of lipid metabolism genes in ex vivo CD4 + T RM cells from CD patients treated with NF-κB inhibitor (BAY/JSH). GAPDH and β-actin served as unmodified negative control, and ICAM-1 was the positive control of NF-kB target gene. ( E ) The mRNA levels of lipid metabolism genes in ex vivo control and RELA/p65 knockdown CD4 + T RM cells from CD patients. GAPDH and β-actin served as unmodified negative control, and ICAM-1 was the positive control of NF-kB target gene. All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with unpaired two-sided t test and Mann-Whitney test in B and one-way analysis of variance in D and E . ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Expressing, Western Blot, Control, MANN-WHITNEY, Immunofluorescence, Staining, Ex Vivo, Negative Control, Positive Control, Knockdown

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Transcription factor NF-κB regulates expression levels of lipid metabolism genes in CD4 + T RM cells. ( A ) Consensus binding motif of RELA/p65 identified in promoter regions of lipid metabolism genes by JASPAR. ( B ) ChIP-qPCR analysis of RELA/p65 binding at promoter regions of lipid metabolism genes in ex vivo CD4 + T RM cells from CD and control patients. ( C ) ChIP-qPCR analysis of RELA/p65 binding at promoter regions of lipid metabolism genes in ex vivo control and RELA/p65 knockdown CD4 + T RM cells from CD patients. GAPDH and β-actin served as unmodified negative control, and ICAM-1 was the positive control of NF-kB target gene. ( D ) Labeling pattern of acetyl-CoA and TCA cycle intermediates (M+2) in isotope tracing ( 13 C 16 -palmitic acid) experiment of ex vivo CD4 + T RM cells from CD patients treated with NF-κB inhibitor (BAY/JSH). All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with one-way analysis of variance in B–D . ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Expressing, Binding Assay, ChIP-qPCR, Ex Vivo, Control, Knockdown, Negative Control, Positive Control, Labeling

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Targeting FAO reverses apoptosis-resistant and proinflammatory phenotype of CD4 + T RM cells in CD patients. ( A ) Apoptosis induced by anti-CD3 in ex vivo CD4 + T RM cells from CD and control patients as determined by flow cytometry. ( B ) Anti-CD3–induced apoptosis rates of ex vivo CD4 + T RM cells from CD and control patients. ( C and D ) Endogenous apoptosis (without anti-CD3 induction) rates in ex vivo CD4 + T RM cells from CD patients treated with FAO inhibitor (etomoxir [ETO], 2 μmol/L; ranolazine [Ran], 5 μmol/L). All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with unpaired two-sided t test in B and one-way analysis of variance in D . ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Ex Vivo, Control, Flow Cytometry

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Targeting FAO reverses apoptosis-resistant and proinflammatory phenotype of CD4 + T RM Cells in CD patients and migration inhibition effect of CD4 + T RM cells on normal intestinal epithelial cell lines. ( A and B ) Anti-CD3–induced apoptosis rates in ex vivo CD4 + T RM cells from CD patients treated with FAO inhibitor (etomoxir, ETO; ranolazine, Rano). ( C and D ) The mRNA and protein levels of indicated cytokines from ex vivo CD4 + T RM cells in CD patients treated with FAO inhibitor. ( E ) Normal intestinal epithelial cell lines were co-cultured with control medium and CM from CD4 + T RM cells treated with vehicle or FAO inhibitors etomoxir in Transwell inserts. Scale bar, 200 μm. ( F ) Quantitative statistical results from Transwell assay. ( G ) Monolayers of normal intestinal epithelial cells were cultured in control medium and CM from CD4 + T RM cells treated with vehicle or FAO inhibitors etomoxir. Scale bar, 400 μm. ( H ) Quantitative statistical results from wound healing assay. All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with one-way analysis of variance. ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Migration, Inhibition, Ex Vivo, Cell Culture, Control, Transwell Assay, Wound Healing Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4 + Tissue-resident Memory T cells in Crohn’s Disease

doi: 10.1016/j.jcmgh.2024.02.014

Figure Lengend Snippet: Targeting FAO reverses migration inhibition effect of CD4 + T RM cells on normal intestinal epithelial cell lines. ( A ) Normal human intestinal epithelial cell lines were co-cultured with control medium and CD4 + T RM cells treated with isotype antibody, anti-TNFα, anti-IL-17, anti-IFN-γ, or their combination in Transwell inserts. Scale bar, 200 μm. ( B and C ) Quantitative statistical results from Transwell assay. ( D and E ) Monolayers of human normal intestinal epithelial cells were co-cultured with control medium and CD4 + T RM cells treated with isotype antibody, anti-TNFα, anti-IL-17, anti-IFN-γ, or their combination. Scale bar, 400 μm. ( F and G ) Quantitative statistical results from wound healing assay. All data were presented as mean values ± SD of minimum of 3 biological replicates. Statistical analysis was performed with one-way analysis of variance in B and C and F and G . ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: CD4 + T RM cells were isolated from the LPMCs by negative selection using human CD4 + T-cell memory effector cell isolation kit (#130-094-125, Miltenyi Biotec).

Techniques: Migration, Inhibition, Cell Culture, Control, Transwell Assay, Wound Healing Assay

Journal: Journal of cell science

Article Title: Rab2a and Rab27a cooperatively regulate the transition from granule maturation to exocytosis through the dual effector Noc2.

doi: 10.1242/jcs.195479

Figure Lengend Snippet: Figure 2. Rab2a interacts with the Noc2-Rab27a binary complex

Article Snippet: The following commercially purchased antibodies were also used: rabbit polyclonal antibodies toward FLAG (F7425, Sigma-Aldrich), hemagglutinin (HA; 561, MBL), green fluorescent protein (GFP; 598, MBL), Rab27a/b (18975, IBL), Noc2 (15297-1-AP, Proteintech), Rab2a (15420-1-AP, Proteintech), ICA69 (ab81500, Abcam), and PICK1 (ab3420, Abcam); and mouse monoclonal antibodies toward Rab3 (610379, BD Biosciences), EEA1 (610457, BD Biosciences), TGN38 (610849, BD Biosciences), PDI (MA3-018, Affinity BioReagents), α-tubulin (T5168, Sigma-Aldrich), and proinsulin (clone 3A1; ab8301, Abcam).

Techniques:

Journal: Journal of cell science

Article Title: Rab2a and Rab27a cooperatively regulate the transition from granule maturation to exocytosis through the dual effector Noc2.

doi: 10.1242/jcs.195479

Figure Lengend Snippet: Figure 3. The N-terminal region of Noc2 is required for binding to Rab2a

Article Snippet: The following commercially purchased antibodies were also used: rabbit polyclonal antibodies toward FLAG (F7425, Sigma-Aldrich), hemagglutinin (HA; 561, MBL), green fluorescent protein (GFP; 598, MBL), Rab27a/b (18975, IBL), Noc2 (15297-1-AP, Proteintech), Rab2a (15420-1-AP, Proteintech), ICA69 (ab81500, Abcam), and PICK1 (ab3420, Abcam); and mouse monoclonal antibodies toward Rab3 (610379, BD Biosciences), EEA1 (610457, BD Biosciences), TGN38 (610849, BD Biosciences), PDI (MA3-018, Affinity BioReagents), α-tubulin (T5168, Sigma-Aldrich), and proinsulin (clone 3A1; ab8301, Abcam).

Techniques: Binding Assay

Journal: Journal of cell science

Article Title: Rab2a and Rab27a cooperatively regulate the transition from granule maturation to exocytosis through the dual effector Noc2.

doi: 10.1242/jcs.195479

Figure Lengend Snippet: Figure 6. Effects of overexpression of Noc2 and its mutants on insulin secretion

Article Snippet: The following commercially purchased antibodies were also used: rabbit polyclonal antibodies toward FLAG (F7425, Sigma-Aldrich), hemagglutinin (HA; 561, MBL), green fluorescent protein (GFP; 598, MBL), Rab27a/b (18975, IBL), Noc2 (15297-1-AP, Proteintech), Rab2a (15420-1-AP, Proteintech), ICA69 (ab81500, Abcam), and PICK1 (ab3420, Abcam); and mouse monoclonal antibodies toward Rab3 (610379, BD Biosciences), EEA1 (610457, BD Biosciences), TGN38 (610849, BD Biosciences), PDI (MA3-018, Affinity BioReagents), α-tubulin (T5168, Sigma-Aldrich), and proinsulin (clone 3A1; ab8301, Abcam).

Techniques: Over Expression

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Elotuzumab, a potential therapeutic humanized anti-SLAMF7 monoclonal antibody, enhances natural killer cell-mediated killing of primary effusion lymphoma cells

doi: 10.1007/s00262-022-03177-6

Figure Lengend Snippet: Elo-mediated ADCC in the FcR engineered Jurkat ADCC indicator system. A BCBL-1 cells were incubated with three different concentrations of Elo, and co-cultured with ADCC-indicating Jurkat cells (E:T ratio = 1:5) for 18 h. Luciferase signal was then measured by luminometer. B GTO and TY-1 cells were incubated with 100 μg/ml of Elo, and co-cultured with ADCC-indicating Jurkat cells (E:T ratio = 1:5) for 18 h. Luciferase signal was then measured by luminometer

Article Snippet: Jurkat T cells expressing firefly luciferase gene under the control of NFAT response elements with constitutive expression of human FcγRIIIa, high affinity (V158) variant and Fcγ chain (ADCC-Jurkat) (#60,541, BPS Bioscience, San Diego, CA) were used to measure ADCC activity by reporter assay.

Techniques: Incubation, Cell Culture, Luciferase

Journal: Experimental and Therapeutic Medicine

Article Title: Management of a rare ovarian carcinosarcoma: A case report and literature review

doi: 10.3892/etm.2022.11520

Figure Lengend Snippet: Immunohistochemical results of bilateral adnexal masses. The results indicated that the serous carcinoma component was (A) positive for cytokeratin and (B) P53, (C) squamous cell carcinoma was positive for P40, and (D) chondrosarcoma was positive for S100 and (E) rhabdomyosarcoma was positive for MyoD1 (magnification, x100).

Article Snippet: Primary antibodies applied in the IHC analysis were mainly as follows: Monoclonal mouse anti-human cytokeratin (CK) (AE1/AE3), mouse anti-human tumor protein p53 monoclonal antibody (DO-7), rabbit polyclonal anti-human S100 protein, monoclonal mouse anti-vimentin (V9; all from Dako; Agilent Technologies, Inc.), mouse anti-human tumor protein P40 monoclonal antibody (cat. no. 66622-1-Ig) and MYOD1 rabbit polyclonal antibody (cat. no. 18943-1-AP; both from ProteinTech Group, Inc.).

Techniques: Immunohistochemical staining