talin  (Santa Cruz Biotechnology)

Journal: The Journal of Biological Chemistry

Article Title: ST3GAL1 regulates cancer cell migration through crosstalk between EGFR and neuropilin-1 signaling

doi: 10.1016/j.jbc.2025.108368

Figure Lengend Snippet: ST3GAL1 regulates cell migration. A , the wound healing ability of MDA-MB-231 cells transfected with control sc or ST3GAL1 siRNA was assessed. B , migration and invasion assays were performed using Transwell inserts, either untreated for the migration assay or coated with Matrigel for the invasion assay. MDA-MB-231 cells transfected with control pVoid or shST3GAL1 were added to the upper chamber of Transwell inserts. The lower chamber was filled with complete culture medium (CM) or 100 ng/ml EGF in serum free media for transwell migration assay. Cells that invaded the inverse side of the membrane were stained with crystal violet and counted. Data are presented as the mean cell numbers/field ± standard deviation, based on representative results from three independent experiments (n = 3). Statistical significance was determined using the two-way ANOVA with multiple comparisons. Scale bar = 100 um. C , total cell lysates from control pVoid- or shST3GAL1 -transfected MDA-MB-231 cells were processed for Western blotting with a talin antibody. The intensity of the cleaved form (short) was first normalized to that of the full-length form (long) and then normalized to the short/long ratio of the control cells. Data are presented as the mean relative ratio ± standard deviation, based on representative results from three independent experiments (n = 3). Statistical significance was determined by two-tailed Student's t test. D , zymography showing changes in calpain activity after ST3GAL1 silencing in MDA-MB-231 cells. MDA-MB-231 ( E ) or MCF7 ( F ) cells transfected with control pVoid or shST3GAL1 were serum starved overnight and then treated with 10 ng/ml EGF for 30 min. CAPN2 was immunoprecipitated from cell lysates and processed for Western blotting with phospho-serine and CAPN2 antibodies. The intensity of phospho-serine was first normalized to that of immunoprecipitated CAPN2 and then normalized to the ratio of phospho-serine/CAPN2 in control cells without EGF treatment. Representative images are shown. Data are presented as the mean relative fold of p-CAPN2/CAPN2 or Talin ratio ± standard deviation, based on representative results from three independent experiments (n = 3). Statistical significance was determined using the two-way ANOVA with multiple comparisons. G , control or ST3GAL1 -silenced MDA-MB-231 cells were treated with 15 ng/ml EGF for 2 h. The transcription of the indicated genes was measured and normalized to that of GAPDH by qPCR analysis. Data are presented as the mean relative fold ± standard deviation, based on representative results from three independent experiments (n = 3). p, control pVoid shRNA; sh, ST3GAL1 shRNA; SF, serum free. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The following antibodies were used in this study: p-AKT-Ser473, AKT, EGFR, NRP1 (Cell Signaling), p-serine (Millipore), p-EGFR, NRP1 (Abcam), EGFR, calpain 2, actin, talin (Santa Cruz), HA (Sigma), GAPDH (Genetex), streptavidin-PE, and alkaline phosphatase-conjugated secondary antibodies (Jackson ImmunoResearch).

Techniques: Migration, Transfection, Control, Invasion Assay, Transwell Migration Assay, Membrane, Staining, Standard Deviation, Western Blot, Two Tailed Test, Zymography, Activity Assay, Immunoprecipitation, shRNA

Journal: Nature Structural & Molecular Biology

Article Title: Talin and kindlin use integrin tail allostery and direct binding to activate integrins

doi: 10.1038/s41594-023-01139-9

Figure Lengend Snippet: Reagents used in the study

Article Snippet: Anti-talin head HRP conjugate (1:100) , Santa Cruz Biotechnology, sc-365875 HRP.

Techniques: Control, DNA Ligation, Labeling, Chromatography, Purification, Cloning

Journal: Nature Structural & Molecular Biology

Article Title: Talin and kindlin use integrin tail allostery and direct binding to activate integrins

doi: 10.1038/s41594-023-01139-9

Figure Lengend Snippet: Reagents used in the study

Article Snippet: Anti-talin head HRP conjugate (1:100) , Santa Cruz Biotechnology, sc-365875 HRP.

Techniques: Control, DNA Ligation, Labeling, Chromatography, Purification, Cloning