(a) Mouse model of MYC induced HCC where MYC is under the control of a tetracycline responsive element (TRE) which contain the tetracycline-controlled transactivator protein (tTA) driven by the …
(a) Representative images from liver and lung (1X and 10X) H and E from liver and lungs of mice with Twist1-transgenic mice demonstrating lack of pathology. (b) Representative images from …
(a) Principal component analysis (PCA) showed that MYC- (n = 5) and MYC/Twist1-HCC (n = 5) overall had distinct, non-overlapping expression profiles. Volcano plot shows comparative analysis of …
(a) The top biological processes upregulated in genes differentially expressed in MYC/Twist1- (n = 5) versus MYC-HCC (n = 5) in transgenic mouse primary tumor tissue. (b) Representative images from …
(a) Schematic of the experiment to extract tumor associated macrophages from the primary tumors of MYC- (n = 5) and MYC/Twist1-HCC (n = 5). Conditioned media from macrophages extracted from MYC-HCC …
(a) Expression of MYC upon Dox treatment of MYC/Twist-HCC cells stably transfected with MYC or Twist1 or control vector, quantified by qPCR. (b) Expression of Twist1 upon Dox treatment of MYC/Twist-H…
(a) Experimental scheme- MYC and MYC/Twist1 cells were implanted orthotopically and metastatic burden in liver and lung assessed after 4 weeks. (b) Representative BLI imaging, gross organ …
(a) Conditional mRNA expression of MYC and Twist1 in primary cell lines derived from MYC-HCC(n = 5, with three technical replicates each) and MYC/Twist1-HCC (n = 5, with three technical replicates …
(a) Experimental scheme- conditioned media (CM) from MYC or MYC/Twist1 cells was used to treat non polarized macrophages for 48 hr. Following that, macrophage migration or polarization was assessed. …
(a) Expression of M1 marker genes in macrophages treated with conditioned media from MYC-HCC (n = 3, with three technical replicates each) or MYC/Twist1-HCC (n = 3, with three technical replicates …
(a) Experimental scheme- Mice orthotopically transplanted with MYC-HCC cells were treated either with PBS or Il13 or Ccl2 or Ccl2+Il13 recombinant cytokines for 4 weeks. (b) MYC expressing single …
(a) Representative images from immunofluorescence staining for Cd206 in liver primary tumor reveals polarization to M2-like phenotype in MYC-HCC bearing mice treated with Il13 and Ccl2+Il13 but not …
(a) Experimental scheme- Mice orthotopically transplanted with MYC/Twist1-HCC cells were treated either with control (ctrl) antibody or anti-Ccl2 antibody (ab) or anti-Il13 ab or anti-Ccl2+Il13 abs …
(a) Representative images from Cd206 staining of liver primary tumor sections (10X) of orthotopic transplanted MYC/Twist1-HCC bearing mice treated with control (ctrl) ab or anti-Ccl2 ab or anti-Il13 …
(a) Disease-free survival in pan-cancer TCGA cohort of 9502 patients from 33 cancers stratified by median MYC+TWIST1 expression (first KM curve) and median MYC+TWIST1+ CCL2+IL13 (second KM curve). (b…
(a) M2 macrophage infiltration in 33 different cancers from the pan-cancer TCGA cohort based on transcriptome deconvolution analysis using CIBERSORT. (b) CIBERSORT analysis of human HCC TCGA RNAseq …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Antibody | MYC (Rabbit, Monoclonal) | Epitomics | RRID:AB_11000313 | IHC (1:150), WB (1:1000) |
Antibody | F4/80 (Rat, Monoclonal) | ThermoFisher | RRID: AB_10376289 | IHC (1:150) |
Antibody | Twist1 (Mouse, Monoclonal) | Abcam | RRID:AB_883294 | WB (1:500) |
Antibody | Phospho-Histone 3 (Rabbit, Polyclonal) | Cell Signaling Technology | RRID:AB_331535 | IHC (1:100) |
Antibody | Cleaved Caspase3 (Rabbit, Polyclonal) | Cell Signaling Technology | RRID:AB_2341188 | IHC (1:100) |
Antibody | Neutrophil (Rat, Monoclonal) | Abcam | RRID:AB_881409 | IHC (1:100) |
Antibody | CD4 (Mouse, Monoclonal) | Abcam | RRID:AB_2686917 | IHC (1:1000) |
Antibody | Glutamine Synthetase (Rabbit, Monoclonal) | Abcam | RRID:AB_446132 | IHC (1:200) |
Antibody | CD31 (Rabbit, Polyclonal) | Abcam | RRID:AB_726362 | IHC(1:100) |
Antibody | CD206 (Mouse, Monoclonal) | R and D | RRID:AB_2745540 | ICC (1:100) |
Antibody | CCL2 (Mouse, Monoclonal) | BioXCell | RRID:AB_10950302 | In vivo treatment (10 mg/kg body weight three times per week) |
Antibody | Il-4 (Mouse, Monoclonal) | Genentech | Propreitary | In vivo treatment (10 mg/kg body weight three times per week) |
Antibody | IL-13 (Mouse, Monoclonal) | Genentech | Propreitary | In vivo treatment (10 mg/kg body weight three times per week) |
Genetic Reagent (M. musculus) | Twist1-tetO7-luc | PMID: 22654667 | Twist1 Transgenic mouse model | Felsher Lab |
Genetic Reagent (M. musculus) | LAP-tTA | PMID: 15475948 | LAP-tTA mouse | Felsher Lab |
Genetic Reagent (M. musculus) | TetO-MYC | PMID: 10488335 | MYC Transgenic mouse | Felsher Lab |
Genetic Reagent (M. musculus) | Nod Scid-Gamma (NSG) mice | Jackson Laboratory | MGI:3577020 | Felsher Lab |
Cytokine | CCL2 | Peprotech | RRID:AB_147738 | In vivo treatment (500 ng/mouse) |
Cytokine | IL13 | Peprotech | RRID:AB_147840 | In vivo treatment (250 ng/mouse) |
Cytokine | IL4 | Peprotech | RRID:AB_147635 | In vivo treatment (250 ng/mouse) |
Drug | Clodronate liposomes | Liposoma | CP-005–005 | In vivo treatment (6.5 μl/g body weight/mouse) |
Sequence-based reagent | MYC | Stanford PAN facility | N/A | f-CTGCGACGAGGAGGAGAACT r-GGCAGCAGCTCGAATTTCTT |
Sequence-based reagent | UBC | Stanford PAN facility | N/A | f-AGCCCAGTGTTACCACCAAG r-ACCCAAGAACAAGCACAAGG |
Sequence-based reagent | Twist1 | Stanford PAN facility | N/A | f-GGACAAGCTGAGCAAGATTCA r-CGGAGAAGGCGTAGCTGAG |
Sequence-based reagent | CD206 | Stanford PAN facility | N/A | f-CTCTGTTCAGCTATTGGACGC r-TGGCACTCCCAAACATAATTTGA |
Sequence-based reagent | Arginase-1 | Stanford PAN facility | N/A | f-CTCCAAGCCAAAGTCCTTAGAG r-AGGAGCTGTCATTAGGGACATC |
Sequence-based reagent | CCR2 | Stanford PAN facility | N/A | f-GTGTACATAGCAACAAGCCTCAAAG r-CCCCCACATAGGGATCATGA |
Sequence-based reagent | INFaR | Stanford PAN facility | N/A | f-CTTCCACAGGATCACTGTGTACCT r-TTCTGCTCTGACCACCTCCC |
Sequence-based reagent | iNOS | Stanford PAN facility | N/A | f-ACACCGACCCGTCCACAGTAT r-CAGAGGGGTAGGCTTGTCTC |
Sequence-based reagent | CX3CR1 | Stanford PAN facility | N/A | f-TACCTTGAGGTTAGTGAACGTCA r-CGCTCTCGTTTTCCCCATAATC |
List of genes (220 up and 294 down) that were differentially expressed between the MYC-HCC and MYC/Twist1-HCC.
The top biological processes, and associated genes, upregulated in MYC/Twist1-HCC versus MYC-HCC.