Journal: Cardiovascular research

Article Title: Functional tissue-engineered blood vessels from bone marrow progenitor cells.

doi: 10.1016/j.cardiores.2007.04.018

Figure Lengend Snippet: Fig. 1. BM-SMPC express biochemical markers of smooth muscle cells. (A) RT-PCR showed that BM-SMPC expressed smooth muscle cell markers as indicated. (B) Western blot for smooth muscle α-actin and calponin; beta-actin served as loading control. (C) Immunocytochemistry for smooth muscle α-actin and calponin. V-SMC from umbilical veins of near-term lambs were used as positive control. Representative results from three independent experiments are shown.

Article Snippet: Western blots were performed as described previously [33,34] using the following antibodies: mouse anti human smooth muscle α-actin (1:100 dilution; SeroTec) and mouse anti-human calponin (1:100 dilution; DakoCytomation) in TBS-Tween.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Control, Immunocytochemistry, Positive Control

Journal: Cardiovascular research

Article Title: Functional tissue-engineered blood vessels from bone marrow progenitor cells.

doi: 10.1016/j.cardiores.2007.04.018

Figure Lengend Snippet: Fig. 2. BM-TEV displayed similar morphologic and biochemical characteristics as TEVs from V-SMC. BM-SMPC or V-SMC were embedded in fibrin hydrogels and cultured around 4-mm mandrel for 2 weeks to form cylindrical tubes. (A) Hematoxylin and eosin (H&E) staining showed that BM-SMPC were distributed uniformly in fibrin hydrogels (bar=100 μm). (B) Immunostaining of BM-TEV and TEV from V-SMC for smooth muscle α-actin and calponin (bar=100 μm).

Article Snippet: Western blots were performed as described previously [33,34] using the following antibodies: mouse anti human smooth muscle α-actin (1:100 dilution; SeroTec) and mouse anti-human calponin (1:100 dilution; DakoCytomation) in TBS-Tween.

Techniques: Cell Culture, Staining, Immunostaining

Journal: Neoplasia (New York, N.Y.)

Article Title: Inhibition of fibronectin accumulation suppresses TUMOR growth

doi: 10.1016/j.neo.2021.06.012

Figure Lengend Snippet: Changes in fibronectin affect blood vessels and proliferation. (A) Tumor lesions exhibit less fibronectin staining in sections from knockdown (Kd) tumors compared to controls (CT). Bars represent 100 μm. (B) Kd tumors contain less fibronectin as determined by ELISA. Pieces obtained from tumor samples were weighed and lysed in protein lysis buffer to evaluate by ELISA. The amount of fibronectin was corrected to total protein measured by the BCA method. N=3/6 replicates. (C) Expression of fibronectin mRNA originating from the cancer cells (human) and the host (murine) was diminished. Using primers specific for human and murine fibronectin it was possible to differentiate between both in qPCR analyses. Human fibronectin was corrected to human HPRT and murine fibronectin to murine β-actin. N=5/3 and 7/5 for the two graphs (left to right). (D) The area of CD31 + blood vessels diminishes as does the number of vessels stained with both CD31 (in red) and the pericyte markers α-smooth muscle actin (α-SMA) or desmin (in green). Examples are shown. Bars represent 100 μm. N=11/10 for CD31 + area and stainings with αSMA and 7/7 for stainings with desmin. (E) Endothelial cells from knockdown tumors produce less fibronectin. Isolated endothelial cells were cultured in the presence of fibronectin-depleted FCS and fibronectin evaluated in the conditioned media by ELISA. N=11/5. (F) Proliferation in tumor sections was diminished in Kd tumors as evidenced by the decrease in the percentage of Ki67 + cells. N=9/9. Examples are shown. Bars represent 100 μm. (G) Tumor sections were stained using TUNEL to mark apoptotic cells. No difference was seen between CT and Kd tumors. N=3/3. Examples are shown. Bars represent 100 μm. All pairs were evaluated using t-test. * P <0.05, ** P <0.005, *** P <0.0001.

Article Snippet: We also used rat anti-mouse CD31 (BIO-RAD; MCA2388GA; 1:100), mouse anti-alpha-smooth muscle actin (α-SMA)-Cy3 (Merck; #A2547; 1:200), rabbit anti-desmin (Dianova; #DLN-13732; 1:100), rabbit monoclonal antibody against YAP (Cell signaling #14074, 1:200), goat anti-rabbit-Alexa 647 (Abcam #ab150079; 1:1000), goat anti-mouse-Cy2 (Dianova; #115-225-166; 1:1000), goat anti-rat-Alexa 594 (Dianova; #112-585-062; 1:1000).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Lysis, Expressing, Isolation, Cell Culture, TUNEL Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: Inhibition of fibronectin accumulation suppresses TUMOR growth

doi: 10.1016/j.neo.2021.06.012

Figure Lengend Snippet: Histology of pUR4-treated tumors confirms changes in matrix and reveals a decrease in proliferation. (A) Stained tumor sections show a decrease in fibronectin and collagen in pUR4-treated animals and no changes in R1R2-treated animals. Bars represent 100 μm. (B) Western blots from the tumors confirm a decrease in fibronectin and collagen with pUR4 treatment, but collagen I did not decrease with R1R2 therapy. N=4/4/4/4 replicates/treatment. Relevant pairs were evaluated by t-tests. * P <0.05. (C) Proliferation was diminished after treatment with pUR4. Sections were stained for Ki67. Bars represent 100 μm. N=9/9/9/9. Pairs were evaluated by t-tests. ** P <0.01. (D) Apoptosis was not affected in vivo by the various treatments. Sections were stained for TUNEL. Examples are shown. Bars represent 100 μm. N=9/9/9/9. Pairs were evaluated by t-tests. (E) Evaluation of blood vessels shows a decrease in area of CD31 + vessels with pUR4 treatment as well as the number of CD31 + αSMA + or CD31 + desmin + double positive vessels (adjusted to the area). Sections from representative tumors with bars representing 100μm. These are intratibial tumors from mice treated for 10 days with the peptides. N=12/12/11/12 for CD31 alone or with α-smooth muscle actin (αSMA) and 11/10/7/7 for staining with desmin. Pairs were compared by t-test. * P <0.05, **** P <0.0001.

Article Snippet: We also used rat anti-mouse CD31 (BIO-RAD; MCA2388GA; 1:100), mouse anti-alpha-smooth muscle actin (α-SMA)-Cy3 (Merck; #A2547; 1:200), rabbit anti-desmin (Dianova; #DLN-13732; 1:100), rabbit monoclonal antibody against YAP (Cell signaling #14074, 1:200), goat anti-rabbit-Alexa 647 (Abcam #ab150079; 1:1000), goat anti-mouse-Cy2 (Dianova; #115-225-166; 1:1000), goat anti-rat-Alexa 594 (Dianova; #112-585-062; 1:1000).

Techniques: Staining, Western Blot, In Vivo, TUNEL Assay