Journal: Experimental and molecular pathology

Article Title: Focal adhesion kinase coordinates costamere-related JNK signaling with muscle fiber transformation after Achilles tenotomy and tendon reconstruction.

doi: 10.1016/j.yexmp.2019.03.006

Figure Lengend Snippet: Fig. 9. Effects of FRNK-mediated inhibition of FAK on hypertrophy signaling. Box Whisker plots of the (specific) phosphorylation of Y397-FAK (A), S2448-mTOR (B), T183/Y185-JNK (C) and S63-cJUN (D) in not tenotomized soleus muscle 7 days after co-transfection of muscle pairs with pCMV & pCMV-FAK and pCMV-FRNK & pCMV-FAK. Circles indicate outliers. + and ++, p < .05 and p < .01 vs. pCMV & pCMV-FAK (repeated-measures ANOVA). Values represented normalized intensities respective to the pCMV & pCMV-FAK transfected muscles. E) Example western blot showing the detection of FAK and FRNK (top) in 10 μg protein homogenate from a soleus muscle pair being co-transfected with pCMV & pCMV-FAK and pCMV-FRNK & pCMV-FAK, respectively. Below, respective actin band.

Article Snippet: An antibody directed against the Nterminal region of FAK (AF4467, R&D systems) and an antibody that detects the phosphorylation at tyrosine 397 (P00151, Boster Biological Technology) were biotinylated (Biotin-NH2 labeling Kit, KT-221, Kamiya Biomedical Company) and used as capture antibodies.

Techniques: Inhibition, Whisker Assay, Phospho-proteomics, Cotransfection, Transfection, Muscles, Western Blot

Journal: Experimental and molecular pathology

Article Title: Focal adhesion kinase coordinates costamere-related JNK signaling with muscle fiber transformation after Achilles tenotomy and tendon reconstruction.

doi: 10.1016/j.yexmp.2019.03.006

Figure Lengend Snippet: Fig. 10. Inter-relationships. Hierarchical networks of the correlations between measured molecular and cellular parameters in all assessed soleus mus- cles. Correlated parameters (nodes) are connected though lines (edges; solid dark-grey line: positive correlation, back- ward slashed grey line: negative correlation). Abbreviations: body_m_e, body mass at the end of the experiment; body_m_s, body mass at the start of the experiment; %con_tis, percentage area of connective tissue; %f-fast, percentage of fast (II) type muscle fibers; %f-hybrid, percentage of hybrid (I/II) type muscle fibers; %f-inuc, percentage of fibers with internal nu- clei; %f-slow, percentage of slow (I) type muscle fibers; FAK, levels of FAK; FRNK, levels of FRNK; g-VN, gamma-vinculin; GM_mass, mass of gastrocnemius muscle; MCSA, mean cross sectional area of muscle fibers; m-VN, meta-vinculin; s-pFAK, specific phosphorylation of Y397-FAK; s-pJNK, specific phosphorylation of T183/Y185-JNK; s-pmTOR, specific phosphorylation of S2448-mTOR; s-pP70S6K, specific phos- phorylation of T421/S424-P70S6K; SOL_mass, mass of soleus muscle.

Article Snippet: An antibody directed against the Nterminal region of FAK (AF4467, R&D systems) and an antibody that detects the phosphorylation at tyrosine 397 (P00151, Boster Biological Technology) were biotinylated (Biotin-NH2 labeling Kit, KT-221, Kamiya Biomedical Company) and used as capture antibodies.

Techniques: Phospho-proteomics

Journal: Biomolecules and Biomedicine

Article Title: KDM3A drives NSCLC proliferation and metastasis via H3K9 demethylation, EMT activation, and MMP-9 upregulation

doi: 10.17305/bb.2025.11251

Figure Lengend Snippet: Knockdown of KDM3A enhances H3K9me2 and modulates the cell invasion signaling pathway. (A) Two siRNAs inhibited KDM3A expression, resulting in an upregulation of H3K9me2 across the three cell lines, with no significant changes observed in KDM3B expression. The upper panel presents representative Western blot images, while the lower panel displays statistical analyses of the Western blot data ( n ═ 3). (B) The expression levels of E-cadherin, vimentin, MMP-9, pFAK, and FAK were evaluated using Western blotting. The upper panel shows representative Western blot images, and the lower panel provides statistical analyses ( n ═ 3). The ratio of pFAK to FAK was calculated after normalization to the β-actin loading control. (C) The expression levels of E-cadherin, vimentin, MMP-9, KDM3B, and KDM3C were assessed using qRT-PCR ( n ═ 3). All experiments were conducted a minimum of three times. Statistical analyses were performed using the Student's t -test, with significance levels indicated as follows: * p < 0.05; ** p < 0.01; *** p < 0.001, ns: no significant differences. Abbreviations: KDM3A: Lysine-specific demethylase 3A; H3K9me2: Histone H3 lysine 9 dimethylation; KDM3B: Lysine-specific demethylase 3B; KDM3C: Lysine-specific demethylase 3C; siRNA: Small interfering RNA; FAK: Focal adhesion kinase; H1299/A549/HCC827: NSCLC cell lines; VIM: Vimentin.

Article Snippet: Membranes were then incubated overnight at 4 ∘ C with antibodies against KDM3A (GeneTex, 54313, 1:1000), KDM3B (Proteintech, 19915-1-AP, 1:1000), H3K9me2 (Cell Signaling Technology, 4658T, 1:2000), β-actin (Santa Cruz Biotechnology, sc-47778, 1:1000), matrix metalloproteinase-9 (MMP-9) (Santa Cruz Biotechnology, sc-393859, 1:1000), E-cadherin (Santa Cruz Biotechnology, sc-8426, 1:1000), Vimentin (Santa Cruz Biotechnology, sc-6260, 1:1000), FAK (Cell Signaling Technology, 3285S, 1:1000), pFAK (Tyr397) (Cell Signaling Technology, 3283, 1:1000), STAT3 (Cell Signaling Technology, 4904, 1:1000), pSTAT3 (Y705) (Cell Signaling Technology, 9131, 1:1000), and GAPDH (Proteintech, HRP-60004, 1:5000).

Techniques: Knockdown, Expressing, Western Blot, Control, Quantitative RT-PCR, Small Interfering RNA

Journal: Cell Death and Differentiation

Article Title: LncRNA CASC9 promotes esophageal squamous cell carcinoma metastasis through upregulating LAMC2 expression by interacting with the CREB-binding protein

doi: 10.1038/s41418-018-0084-9

Figure Lengend Snippet: CASC9 activates FAK-PI3K/Akt signaling pathways through LAMC2. a Western blot analysis showed altered levels of pFAK, pPI3K, and pAkt after CASC9 or LAMC2 knockdown in KYSE150 and KYSE450 cells. b Western blot analysis of FAK-PI3K/Akt signaling pathways of the rescue experiment. c RT-qPCR analysis of MMP10 and MMP13 expression. d , e The expression of MMP10 and MMP13 in KYSE150 and KYSE450 cells after CASC9 knockdown. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: An anti-GAPDH antibody (#KC-5G5, KangChen, China) was used as an internal reference; anti-Laminin-5 (γ2 chain) (MAB19562) was from Millipore (Millipore, USA); anti-histone H3 (D2B12), anti-ITGB4 (#4707), FAK Antibody Sampler Kit (#9330), anti-PI3K Kinase p110α (#4249), anti-Phospho-PI3 Kinase p85 (Tyr458)/p55(Tyr199) (#4228), anti-AKT (pan) (#4691), and anti-phospho-Akt (Ser473) (#4060) were from Cell Signaling Technology (CST, USA); anti-MMP10 antibody (ab199688) and anti-MMP13 (ab39021), antibody anti-KAT3A/CBP (ab2832), and anti-histone H3 (acetyl K27) (ab4729) were from Abcam (Abcam, UK).

Techniques: Protein-Protein interactions, Western Blot, Knockdown, Quantitative RT-PCR, Expressing

Journal: Cell Death and Differentiation

Article Title: LncRNA CASC9 promotes esophageal squamous cell carcinoma metastasis through upregulating LAMC2 expression by interacting with the CREB-binding protein

doi: 10.1038/s41418-018-0084-9

Figure Lengend Snippet: Model of CASC9 interaction with CBP and the signaling pathways involved in pro-metastatic function. a In the nucleus, CASC9 recruits CBP and modulates H3K27ac levels at the LAMC2 promoter, thereby facilitating gene transcription. b In the cytoplasm, LAMC2 is exported to the extracellular matrix and interacts with ITGB4 to activate FAK-PI3K/Akt signaling pathways, thereby promoting ESCC metastasis

Article Snippet: An anti-GAPDH antibody (#KC-5G5, KangChen, China) was used as an internal reference; anti-Laminin-5 (γ2 chain) (MAB19562) was from Millipore (Millipore, USA); anti-histone H3 (D2B12), anti-ITGB4 (#4707), FAK Antibody Sampler Kit (#9330), anti-PI3K Kinase p110α (#4249), anti-Phospho-PI3 Kinase p85 (Tyr458)/p55(Tyr199) (#4228), anti-AKT (pan) (#4691), and anti-phospho-Akt (Ser473) (#4060) were from Cell Signaling Technology (CST, USA); anti-MMP10 antibody (ab199688) and anti-MMP13 (ab39021), antibody anti-KAT3A/CBP (ab2832), and anti-histone H3 (acetyl K27) (ab4729) were from Abcam (Abcam, UK).

Techniques: Protein-Protein interactions

Journal: Environmental toxicology

Article Title: Gallic acid attenuates metastatic potential of human colorectal cancer cells through the miR-1247-3p-modulated integrin/FAK axis.

doi: 10.1002/tox.24087

Figure Lengend Snippet: FIGURE 5 Involvement of miR-1247-3p in the Gallic acid (GA)-inhibited integrin/FAK cascade in DLD-1 cells. Cells were treated with 90 μM GA for 24 h, lysed for total RNA extraction, and subjected to (A) miRNA expression profiling assay via microarray analysis or (B) quantitative analysis of the miRNAs via qPCR. miRNAs with up- and downregulation in response to GA were labeled with red and green, respectively. (C) Cells were transfected with control vector (Ctl-miR) or anti-miR-1247-3p (anti-miR), treated with GA for 24 h, and subjected to the assessment of integrins, FAK, and paxillin by Western blot. Densitometric analysis was performed for semi-quantitation of signals. The tubulin signal was used as internal control. Average signal ratios of phosphorylated protein/total protein compared with the control were indicated.

Article Snippet: Antibodies specifically recognizing human tubulin (SC-134237), integrin αV (SC-376156), integrin β3(SC-365679), focal adhesion kinase (FAK, SC-271126), phosphorylated FAK (pY397-FAK, SC-81493), paxillin (SC-136297), phosphorylated paxillin (pY118-Paxillin, SC-365020), Src (SC-32789), phosphorylated Src (pY416-Src, SC-24621, and CS#2101), and peroxidase-conjugated antibodies against mouse IgG or rabbit IgG were obtained from Santa Cruz Biotechnology, Inc. (CA, USA).

Techniques: RNA Extraction, Expressing, Microarray, Labeling, Transfection, Control, Plasmid Preparation, Western Blot, Quantitation Assay

Journal: Environmental toxicology

Article Title: Gallic acid attenuates metastatic potential of human colorectal cancer cells through the miR-1247-3p-modulated integrin/FAK axis.

doi: 10.1002/tox.24087

Figure Lengend Snippet: FIGURE 4 Gallic acid (GA) reduced integrin expression and inhibited FAK/Paxillin/Src and PI3K/AKT signaling in DLD-1 cells. Cells were treated with GA at the indicated concentrations for 24 h; collected; and lysed for the immunodetection of (A) integrins and the associated signaling proteins, (B) phosphor-paxillin and Src, and (C) PI3K, phosphor-AKT, and AKT via Western blot. Semi-quantitation of signals was conducted by densitometric analysis. DMSO treatment was used as control (C). Densitometric analysis was performed for the semi-quantitation of signals. The tubulin signal was used as internal control. Average signal ratios of phosphorylated protein/total protein compared with the control were indicated.

Article Snippet: Antibodies specifically recognizing human tubulin (SC-134237), integrin αV (SC-376156), integrin β3(SC-365679), focal adhesion kinase (FAK, SC-271126), phosphorylated FAK (pY397-FAK, SC-81493), paxillin (SC-136297), phosphorylated paxillin (pY118-Paxillin, SC-365020), Src (SC-32789), phosphorylated Src (pY416-Src, SC-24621, and CS#2101), and peroxidase-conjugated antibodies against mouse IgG or rabbit IgG were obtained from Santa Cruz Biotechnology, Inc. (CA, USA).

Techniques: Expressing, Immunodetection, Western Blot, Quantitation Assay, Control

Journal: Artificial cells, nanomedicine, and biotechnology

Article Title: Low intensity pulsed ultrasound promotes the migration of bone marrow- derived mesenchymal stem cells via activating FAK-ERK1/2 signalling pathway.

doi: 10.1080/21691401.2019.1657878

Figure Lengend Snippet: Figure 4. Western blot analysis was in BMSCs induced using LIPUS. (A) BMSCs were stimulated using LIPUS and collected for Western blot analysis at the indicated time point after LIPUS treatment. (B–D) Phosphorylation of FAK, ERK1/2 and MLC induced using LIPUS were analyzed using Western blot in the presence of the spe- cific inhibitors of FAK (PF-573228) and ERK1/2 (U0126).

Article Snippet: After washing, the membrane was blocked with 5% nonfat milk for 1 h, and then incubated overnight at 4 C with specific primary antibodies for FAK, phospho-FAK (Tyr397) (all from Novus, USA), ERK1/2, phospho-ERK1/2, phospho-myosin light chain 2 (ser19) (all from Cell Signalling Technology, USA).

Techniques: Western Blot, Phospho-proteomics