Journal: QJM : monthly journal of the Association of Physicians

Article Title: Snail1-dependent cancer-associated fibroblasts induce epithelial-mesenchymal transition in lung cancer cells via exosomes.

doi: 10.1093/qjmed/hcz093

Figure Lengend Snippet: Figure 2. CAFs induce lung cancer cell line EMT. The protein expression levels of E-cadherin, Vimentin, ZEB1, Snail1 and Twist1 in the lung cancer cell lines co-cultured with CAFs were measured by immunoblotting, using GAPDH protein as the loading control.

Article Snippet: Primary antibodies against the following were used: E-cadherin (ab76055, 1:1000; Abcam, Cambridge, MA, USA), vimentin (D21H3, 1:1000; Cell Signaling Technology, Danvers, MA, USA), ZEB1 (NBP2-23484; 1:800, Novus Biologicals, Littleton, CO, USA), Twist1 (ab175430, 1:1000, Abcam), Snail1(C15D3, 1:1000; Cell Signaling Technology, Danvers, MA, USA), CD81 (ab79559, 1:1000; Abcam, Cambridge, MA, USA) and GAPDH (14C10, 1:1000; Cell Signaling Technology).

Techniques: Expressing, Cell Culture, Western Blot, Control

Journal: iScience

Article Title: Efficient genome editing in erythroid cells unveils novel MYB target genes and regulatory functions

doi: 10.1016/j.isci.2023.107641

Figure Lengend Snippet:

Article Snippet: rabbit anti-Zeb1 , Novus , Cat# NBP2-20942.

Techniques: Recombinant, Membrane, Magnetic Beads, Produced, Protease Inhibitor, DNA Extraction, Isolation, Software

Journal: Oncogenesis

Article Title: miR-200b restoration and DNA methyltransferase inhibitor block lung metastasis of mesenchymal-phenotype hepatocellular carcinoma

doi: 10.1038/oncsis.2012.15

Figure Lengend Snippet: Transient delivery of pre-miR-200a or pre-miR-200b in mesenchymal cells partially reversed EMT. ( a ) Mesenchymal (M) cells demonstrate loss of four miR-200 family members including miR-200a, miR-200b, miR-200c and miR-429 compared with parent epithelial (E) cells ( n =3, * P <0.05). ( b ) Zeb1 and Zeb2 were significantly downregulated and E-cadherin was significantly upregulated in precursor miR-200-transfected mesenchymal cells compared with cells transfected with NC pre-miRNA ( n =3, * P <0.05). ( c ) Protein expression of E-cadherin was significantly upregulated after miR-200 transfection in mesenchymal cells. ( d ) Compared with NC miRNA, the transient expression of miR-200a or miR-200b significantly inhibited tumor cell migration on wound-healing assay ( n =3, NC 149±21, miR-200a 54±10 and miR-200b 26±8, * P <0.05 compared with NC).

Article Snippet: The polyclonal E-cadherin antibody for western blot, immunohistochemistry and immunofluorescence, and polyclonal Zeb1 antibody for western blot were purchased from Cell Signaling (Cat# 3195s and 3396s, respectively, Danvers, MA, USA), and Zeb1 antibody for immunohistochemistry and Zeb2 antibody for immunohistochemistry and western blot were purchased from Novus Biologicals (Littleton, CO, USA) with catalog numbers NBP1-05987 and NBP1-67782, respectively.

Techniques: Transfection, Expressing, Migration, Wound Healing Assay

Journal: Oncogenesis

Article Title: miR-200b restoration and DNA methyltransferase inhibitor block lung metastasis of mesenchymal-phenotype hepatocellular carcinoma

doi: 10.1038/oncsis.2012.15

Figure Lengend Snippet: Stable expression of miR-200b reverses EMT. ( a ) In mesenchymal cells, miR-200b expression results in downregulation of Zeb1 and Zeb2 , and upregulation of E-cadherin compared with NC miRNA (miR-NC, n =3/group, * P <0.01). ( b ) The protein level of E-cadherin was upregulated in miR-200b expressing mesenchymal cells compared with control miR-NC cells, but remained below the level of parent epithelial cells ( n =3, representative image shown). ( c ) Green fluorescence protein (GFP) fluorescence demonstrates that mesenchymal cells undergo morphology change to larger cells with more cell–cell contact after miR-200b upregulation ( n =10, representative image shown). ( d ) Compared with miR-NC control mesenchymal cells, invasion is significantly inhibited after miR-200b expression (expressed as mean number of cells invading into lower chamber, n =3, * P <0.05). ( e ) Mesenchymal cells expressing miR-NC or miR-200b spread to multiple abdominal organs following orthotopic transplant in nude mice, as demonstrated by in vivo imaging ( n =8–9 mice/group, representative images shown). ( f ) Histology demonstrates mesenchymal cells with miR-200b expression invade the pancreas (P) and colon (C) (L, liver; T, tumor). Metastatic lesions (arrow) are detectable in the lungs of both miR-NC and miR-200b groups (miR-NC, n =9, lung metastasis 22.2% (2/9), miR-200b, n =8, lung metastasis 37.5% (3/8), H&E stain).

Article Snippet: The polyclonal E-cadherin antibody for western blot, immunohistochemistry and immunofluorescence, and polyclonal Zeb1 antibody for western blot were purchased from Cell Signaling (Cat# 3195s and 3396s, respectively, Danvers, MA, USA), and Zeb1 antibody for immunohistochemistry and Zeb2 antibody for immunohistochemistry and western blot were purchased from Novus Biologicals (Littleton, CO, USA) with catalog numbers NBP1-05987 and NBP1-67782, respectively.

Techniques: Expressing, Control, Fluorescence, In Vivo Imaging, Staining