bs 3159r  (Bioss)

Journal: APL Bioengineering

Article Title: Survivin modulates stiffness-induced vascular smooth muscle cell motility

doi: 10.1063/5.0252766

Figure Lengend Snippet: Survivin regulates FAK phosphorylation, actin organization, and stress fiber formation. (a) VMSCs were sparsely seeded on fibronectin-coated soft and stiff hydrogels and treated with either YM155 or DMSO. Cells were stained with DAPI, phospho-FAK (Tyr397) antibody and Alexa Fluor 647-phalloidin. Fluorescence images were acquired using a spinning disk confocal microscope with a 63× oil-immersion objective. The outlined boxes indicate the regions shown in the magnified insets. (b) A sum of slices projection of each cell was generated using Fiji/ImageJ, and FAK pY397 clusters were manually counted. 37 cells (soft-DMSO), 44 cells (stiff-DMSO), 33 cells (stiff-0.1 µM YM155), and 37 cells (stiff-2µM YM155) were analyzed from n = 3 independent biological replicates. * p < 0.05, ** p < 0.01.

Article Snippet: For phospho-FAK clustering assay, the same experimental procedures and conditions were used as described above, except cells were stained with phospho-FAK (Tyr397) antibody (1:25; Cat. No. 3283, Cell Signaling Technology) and Alexa Fluor 647-phalloidin (1:200; Cat. No. A22287, Thermo Fisher Scientific).

Techniques: Phospho-proteomics, Staining, Fluorescence, Microscopy, Generated

Journal: The FEBS journal

Article Title: Cadherin-6 controls neuronal migration during mouse neocortical development via an integrin-mediated pathway.

doi: 10.1111/febs.70150

Figure Lengend Snippet: Fig. 7. KD and overexpression of Cdh6 did not alter the levels of Paxillin protein and phosphorylated AKT. (A) Immunostaining for Paxillin (top, red) or pFAK (red, bottom) and EGFP (green) using sections from the E18.0 cerebral cortex, which were electroporated with control (n = 3) or Cdh6 KD vector (n = 3) (left) or control (CAG empty) (n = 3) or CAG-Cdh6-HA vectors (n = 3) (right) with CAG-EGFP vectors at E14.0. (B) Western blot analysis of neocortical cells electroporated with CAG empty or CAG-Cdh6- HA vectors using antibodies against HA, Paxillin, pFAK, FAK, and GAPDH (a loading control) (n = 3). Sections in (A) were stained with DAPI (magenta). Scale bar: 50 lm in (A). KD, knockdown.

Article Snippet: The primary antibodies used in this study were as follows: CDH6 (sheep; R&D Systems, Minneapolis, MN, USA, AF2715, 1 : 200), HA (rat; Roche, Basel, Switzerland, 3F10, 1 : 500), Nestin (chick; Aves Labs, Davis, CA, USA, #NES, 1 : 400), DCX (rabbit; CST, Danvers, MA, USA, #4604, 1 : 500), RORB (mouse; Perseus Proteomics, Tokyo, Japan, PPN7927-00, 1 : 400), CUX1 (rabbit; Proteintech, Rosemont, IL, USA, 11733-1AP, 1 : 600), CD29 (clone 9EG7) (rat; BD, San Jose, CA, USA, 550531), Paxillin (rabbit; Abcam, Cambridge, MA, USA, ab32084, 1 : 200), pFAK (Y397) (rabbit, CST, 3283S, 1 : 1000).

Techniques: Over Expression, Immunostaining, Control, Plasmid Preparation, Western Blot, Staining, Knockdown