Journal: American Journal of Cancer Research

Article Title: Multi-organ metastasis as destination for breast cancer cells guided by biomechanical architecture

doi:

Figure Lengend Snippet: Compared to empty vector, WT-ASPH significantly enhanced metastatic capability of MDA-MB-231 cells, which was efficiently reversed by the SMI in experimental pulmonary metastatic (tail vein injection) murine model. (A) Experimental design and (B) Therapeutic protocol for tail vein injection model (n = 5/group). (C) Using fluorescent imaging system to detect potential pulmonary metastasis in mice from different groups of tail vein injection model. (D) The number of macro-metastases in the lungs derived from mice in tail vein injection model. *P < 0.05; **P < 0.01. (E) Gross appearance of the lungs derived from representative mice in tail vein injection model. Metastatic lesions were highlighted with yellow arrows. (F) Gross appearance and histopathologic characteristics of (Upper) hepatic and (Bottom) pulmonary metastatic lesions of a representative mouse in ASPH+DMSO group of tail vein injection model. Noted the metastatic lesions also maintain high expression of ASPH. This animal was euthanized at 7th weeks. (G) Gross appearance of bone (spine) metastasis derived from a representative mouse in tail vein injection model. The mouse was tail vein injected with ASPH overexpressing MDA-MB-231 cells and treated with DMSO. (H) Histologic characteristics of bone and lung lesions in this specific mouse in (P). (I, J) Expression profiling of key components in SRC signaling pathway (p-SRC Y416, SRC regulator ADAM12 and downstream MMPs) was substantially downregulated by SMI.

Article Snippet: As previously described [ 22 - 24 ], IHC was performed on 4-μm thick FFPE slides with antibodies to ASPH (FB50, homemade); MMP1 (ab38929), MMP14 (ab3644) from Abcam (Cambridge, MA); SRC Y416 ( {"type":"entrez-protein","attrs":{"text":"PAB25310","term_id":"1236638637","term_text":"PAB25310"}} PAB25310 ) from Abnova (Taipei, Taiwan) and ADAM12 (14139-1-AP) from Proteintech (Rosemont, IL).

Techniques: Plasmid Preparation, Injection, Imaging, Derivative Assay, Expressing

Journal: Journal of Cell Science

Article Title: A unique role for clathrin light chain A in cell spreading and migration

doi: 10.1242/jcs.224030

Figure Lengend Snippet: CLCa depletion significantly inhibits spreading-induced signaling. (A) siRNA-transfected cells held in suspension for 1 h (left panel) or plated on collagen IV-coated dishes for the indicated times (minutes, right panel) were lysed and subjected to western blotting with anti-active FAK [pFAK(Y397)], active Src [pSrc(Y416)] and anti-phosphorylated paxillin [pPax(Y118)] antibodies. (B) Protein phosphorylation in control cells at 30 min after plating was set as 100%. The results represent a summary from five to seven experiments. *P<0.05; **P<0.01. (C) Lysates from plated cells treated as in A were analyzed by western blotting with antibodies against Src-dependent FAK phosphorylation sites (Y576 and Y925). The blots shown represent one of three independent experiments.

Article Snippet: Antibodies against the following proteins were used: CLCa (1:1000, sc-28276), CLCb (1:500, sc-376414), actin (1:1000, sc-1616) from Santa Cruz Biotechnology, FAK (1:2000, 610088) and β1-integrin (1:1000, 610467) from BD Transduction Labs, phosphorylated FAK(Y397) (1:1000, 44-624G), phosphorylated paxillin(Y118) (1:1000, 44-722G) from Fisher Scientific, Src (1:2000, 2108), phosphorylated Src(Y416) (1:1000, MAB2685, 2101), phosphorylated FAK(Y576) (1:1000, 3281), FAK(Y925) (1:1000, 3284) from Cell Signaling, phosphorylated Src(Y416) (1:1000, MAB2685) from RD Systems, WAVE1/Scar (1:1000, 07-037), Rac1 (1:2000, 05-389) from Millipore.

Techniques: Transfection, Western Blot