Journal: Frontiers in Oncology

Article Title: Dissection of the signal transduction machinery responsible for the lysyl oxidase-like 4-mediated increase in invasive motility in triple-negative breast cancer cells: mechanistic insight into the integrin-β1-NF-κB-MMP9 axis

doi: 10.3389/fonc.2024.1371307

Figure Lengend Snippet: Identifying TRAF4 and TAK1 on the integrin-β1 signal cascade that links to NF-κB activation. (A) Schematic representation of the NF-κB activation flow via canonical and noncanonical pathways. (B, C) As indicated, the prepared cell lysates of HEK293T transfectants with different combinations of the plasmids were divided into three aliquots. The first was used as the whole cell lysate, i.e., as input (rightmost). The second was used to conduct immunoprecipitations with anti-HA tag antibody-conjugated agarose beads (middle). The remaining third was used to perform similar immunoprecipitations with anti-DYKDDDDK tag antibody-conjugated agarose beads as a negative control for the immunoprecipitation experiment (leftmost). After immunoprecipitation, the expressed products with beads-bound foreign proteins were analyzed by WB with an anti-HA, anti-Myc, or anti-albumin antibody. (D) The indicated cell extracts were incubated with the biotinylated (bio)-integrin-β1 antibody to capture the intrinsic integrin-β1 of the individual cells. On the other hand, bio-control IgG was used as a negative control for the same immunoprecipitation experiment. The captured integrin-β1 was collected by the pull-down method with avidin beads, and the precipitates were further subjected to WB with the indicated antibodies.

Article Snippet: The antibodies used were as follows: mouse anti-HA tag antibody (clone 6E2; Cell Signaling Technology, Danvers, MA), mouse anti-Myc tag antibody (clone 9B11; Cell Signaling Technology), rabbit anti-human MMP9 antibody (Cell Signaling Technology), rabbit anti-human Akt antibody (Cell Signaling Technology), rabbit anti-human phospho (T308)-Akt antibody (Cell Signaling Technology), rabbit anti-human phospho (S473)-Akt antibody (Cell Signaling Technology), rabbit anti-human NF-κB antibody (Cell Signaling Technology), mouse anti-human phospho (S536)-NF-κB antibody (Cell Signaling Technology), rabbit anti-human IKKα antibody (Cell Signaling Technology), rabbit anti-human IKKβ antibody (Cell Signaling Technology), rabbit anti-human phospho (S176/180)-IKKα/β antibody (Cell Signaling Technology), mouse anti-human Iκβα antibody (Cell Signaling Technology), rabbit anti-human phospho (S32)-Iκβα antibody (Cell Signaling Technology), rabbit anti-human integrin-β1 antibody (Proteintech), rabbit anti-human TRAF4 antibody (Proteintech), rabbit anti-human TAK1 antibody (Cell Signaling Technology), rabbit anti-DsRed (RFP) (Takara Bio USA, Mountain View, CA), rabbit anti-GFP (Thermo Fisher Scientific), mouse anti-β-actin (Merk Sigma-Aldrich), and rabbit anti-human albumin (Agilent’s DAKO, Glostrup, Denmark) that cross-reacts with bovine albumin with high affinity in WB.

Techniques: Activation Assay, Negative Control, Immunoprecipitation, Incubation, Avidin-Biotin Assay

Journal: Turkish Journal of Obstetrics and Gynecology

Article Title: Evaluation of endometrial receptivity in recurrent pregnancy loss and recurrent implantation failure

doi: 10.4274/tjod.galenos.2024.42959

Figure Lengend Snippet: Staining changes of the endometrial β1 integrin, FAK, HOXA-11, CD44, and ECM1 in the RPL and RIF groups compared to the control group

Article Snippet: The antibodies used were polyclonal rabbit anti-human against HOXA-11 (1:500 dilutions, Thermo Fischer Scientific, US), monoclonal rabbit anti-human against β1 integrin (clone: EPR16895, 1:1000 dilutions, Abcam, US), monoclonal rabbit anti-human against FAK (clone: EP69Y, 1:250 dilutions, Abcam, US), monoclonal rabbit anti-human against CD44 (clone: EPR1013Y, 1:100 dilutions, Abcam, US), and monoclonal rabbit anti-human against Extracellular Matrix Protein-1 (clone: EPR6701, 1:250 dilutions, Abcam, US).

Techniques: Staining

Journal: Turkish Journal of Obstetrics and Gynecology

Article Title: Evaluation of endometrial receptivity in recurrent pregnancy loss and recurrent implantation failure

doi: 10.4274/tjod.galenos.2024.42959

Figure Lengend Snippet: Endometrial staining in all groups When the pictures were examined in order, β1 Integrin had intense glandular staining in all groups. Although stromal staining was strong in the control and RIF groups, stromal staining was not observed in the RPL group. What is remarkable for FAK is the absence of glandular staining in the RIF group. CD44 did not show stromal staining in the RIF group; in the RPL group, neither stromal nor glandular staining was observed. The absence of glandular ECM1 staining was noted in the RIF group. RPL: Recurrent pregnancy loss, RIF: Recurrent implantation failure, FAK: Focal adhesion kinase, ECM1: Extracellular matrix protein 1 CD44: Cluster of differentiation 44

Article Snippet: The antibodies used were polyclonal rabbit anti-human against HOXA-11 (1:500 dilutions, Thermo Fischer Scientific, US), monoclonal rabbit anti-human against β1 integrin (clone: EPR16895, 1:1000 dilutions, Abcam, US), monoclonal rabbit anti-human against FAK (clone: EP69Y, 1:250 dilutions, Abcam, US), monoclonal rabbit anti-human against CD44 (clone: EPR1013Y, 1:100 dilutions, Abcam, US), and monoclonal rabbit anti-human against Extracellular Matrix Protein-1 (clone: EPR6701, 1:250 dilutions, Abcam, US).

Techniques: Staining