Journal: The Journal of Cell Biology

Article Title: uPAR promotes formation of the p130Cas–Crk complex to activate Rac through DOCK180

doi: 10.1083/jcb.200712050

Figure Lengend Snippet: DOCK180 is required for uPAR-driven membrane ruffling and Rac activation. (A) BE colon carcinoma cells transfected with indicated siRNAs were plated on vitronectin-coated coverslips for 12 h, fixed, and stained with Texas red–conjugated phalloidin. Bar, 50 μm. (B and C) HEK 293T cells were transfected with siRNAs targeting DOCK180 or nontargeting control (NT), and 48 h later were transfected with uPAR expression vector or empty vector control. After 24 h, Rac-GTP pull-down assays were performed. (B) Representative immunoblots to show pull-down assay, uPAR expression, and DOCK180 silencing. (C) Rac activation was quantitated and analyzed as described in Materials and methods (mean + SEM; n = 3). *, P < 0.05; **, P < 0.01; unpaired Student's t test.

Article Snippet: Controls used were AllStars nontargeting control (QIAGEN), ON-TARGET nontargeting control SMART Pool (Thermo Fisher Scientific), and nontargeting control SMART Pool (Thermo Fisher Scientific).

Techniques: Activation Assay, Transfection, Staining, Expressing, Plasmid Preparation, Western Blot, Pull Down Assay

Journal: The Journal of Cell Biology

Article Title: uPAR promotes formation of the p130Cas–Crk complex to activate Rac through DOCK180

doi: 10.1083/jcb.200712050

Figure Lengend Snippet: DOCK180 is required for Rac activation and invasion in uPAR-expressing tumor cell lines. (A) BE (closed bars), MDA-MB-231 (open bars), and SNB19 (shaded bars) cells were transfected with siRNAs. NT, nontargeting control; NT-OT, ON-TARGET nontargeting control. Rac activity was quantitated at 72 h (mean + SEM; n ≥ 3). *, P < 0.05; **, P < 0.01; unpaired Student's t test. (inset) Representative immunoblots from one Rac pull down in BE cells. Irrelevant lanes were removed (represented by vertical black lines). (B) BE (closed bars) and MDA-MB-231 (open bars) cells transfected with siRNAs were assayed for collagen-I invasion (mean + SEM; n ≥ 3). *, P < 0.05; **, P < 0.01; unpaired Student's t test. Immunoblots showing knockdown are in Fig. S2 A, available at http://www.jcb.org/cgi/content/full/jcb.200712050/DC1 .

Article Snippet: Controls used were AllStars nontargeting control (QIAGEN), ON-TARGET nontargeting control SMART Pool (Thermo Fisher Scientific), and nontargeting control SMART Pool (Thermo Fisher Scientific).

Techniques: Activation Assay, Expressing, Transfection, Activity Assay, Western Blot

Journal: The Journal of Cell Biology

Article Title: uPAR promotes formation of the p130Cas–Crk complex to activate Rac through DOCK180

doi: 10.1083/jcb.200712050

Figure Lengend Snippet: β 3 integrin is required for Rac activation in uPAR-expressing cells. (A) Tumor cells were transfected with siRNAs and Rac activation was quantitated. Closed bars, BE; open bars, MDA-MB-231; shaded bars, SNB19 (mean + SEM; n ≥ 4). *, P < 0.05; **, P < 0.01; unpaired Student's t test. (B) BE (closed bars) and MDA-MB-231 cells (open bars) were transfected with siRNAs and assayed for collagen-I invasion (mean + SEM; n ≥ 4). *, P < 0.05; **, P < 0.01; unpaired Student's t test. (C) BE cells were seeded for invasion assays in serum-free DME or DME + 5% serum and either control IgG or antivitronectin antibody (mean + SEM; n ≥ 4). **, P < 0.01; unpaired Student's t test. (D) HEK 293T cells were transfected with siRNAs, and after 48 h, transfected with uPAR or empty vector. Rac activity was measured 24 h later. (E) Quantitation of Rac-activation assays in HEK 293T cells (mean + SEM; n ≥ 5). *, P < 0.05; unpaired Student's t test versus nontargeting. Immunoblots showing knockdown are in Fig. S2 (C and D), available at http://www.jcb.org/cgi/content/full/jcb.200712050/DC1 .

Article Snippet: Controls used were AllStars nontargeting control (QIAGEN), ON-TARGET nontargeting control SMART Pool (Thermo Fisher Scientific), and nontargeting control SMART Pool (Thermo Fisher Scientific).

Techniques: Activation Assay, Expressing, Transfection, Plasmid Preparation, Activity Assay, Quantitation Assay, Western Blot

Journal: Cell Death & Disease

Article Title: Regulation of senescence escape by TSP1 and CD47 following chemotherapy treatment

doi: 10.1038/s41419-019-1406-7

Figure Lengend Snippet:

Article Snippet: pLKO , Nontarget shRNA control , MISSION shRNA Sigma SHC216 , Nontarget shRNA control.

Techniques: Plasmid Preparation, Sequencing, shRNA

Journal: Communications Biology

Article Title: β-arrestin mediates communication between plasma membrane and intracellular GPCRs to regulate signaling

doi: 10.1038/s42003-020-01510-2

Figure Lengend Snippet: shRNA sequences.

Article Snippet: Scramble (nontargeting) , Sigma-SCH002.

Techniques: shRNA, Plasmid Preparation, Sequencing