Journal: The Journal of Pathology

Article Title: Lactate‐mediated activation of GPR81 regulates BCR /Abl protein expression in chronic myeloid leukemia cells selected under low oxygen tension

doi: 10.1002/path.6492

Figure Lengend Snippet: GPR81 activation drives lactate‐mediated suppression of BCR/Abl protein expression and signaling and its effects on the maintenance of stem cell potential in low oxygen. K562 or KCL22 cells were incubated at 3 × 10 5 cells/ml in atmosphere at 0.1% O 2 (LC1) for 4 or 7 days, respectively, in the presence or absence of different concentrations of (A) the GPR81 antagonist 3‐OBA (1–10 m m ) or (B) the lactate transporter inhibitor syrosingopine (10 μ m ) and/or the GPR81 agonist 3‐chloro‐5‐hydroxybenzoic acid (GPR81ago). (A and B) Total cell lysates were subjected to SDS‐PAGE and immunoblotting with anti‐c‐Abl or anti‐p‐Crkl Ab; anti‐vinculin Ab was used to verify equal protein loading. One representative experiment out of three with similar results is shown. (C) Cells rescued at end of LC1 were replated at 3 × 10 4 cells/ml and incubated at 21% O 2 (LC2). Trypan blue‐negative cells were counted at the times of incubation in LC2 indicated in abscissa. Values are mean ± SD from three independent experiments; * p < 0.05 versus control; † p < 0.05 versus syrosingopine.

Article Snippet: The primary antibodies used were c‐Abl (K‐12), rabbit polyclonal; phospho‐Crkl (Tyr207), rabbit polyclonal (Cell Signaling Technology, Danvers, MA, USA); Gpr81 (Catalogue No.: 58‐342, ProSci LLC, California, USA), rabbit polyclonal; vinculin (clone hVIN‐1), mouse monoclonal (Sigma‐Aldrich).

Techniques: Activation Assay, Expressing, Incubation, SDS Page, Western Blot, Control

Journal: international Journal of Oncology

Article Title: Expression and potential function of the CXC chemokine CXCL16 in pancreatic ductal adenocarcinoma

doi: 10.3892/ijo_00000009

Figure Lengend Snippet: Figure 1. Expression of CXCL16 and CXCR6 mRNA in pancreatic cancer (PDAC) cells and pancreatic tissues (qRT-PCR). (A) CXCL16 and (B) CXCR6 mRNA expression levels in pancreatic cancer cell lines. (C) CXCL16 mRNA expression levels in normal pancreas (n=10), chronic pancreatitis (CP) (n=11), and PDAC tissue (n=11); (P<0.001 PDAC versus normal, P<0.001 PDAC versus CP). (D) CXCR6 mRNA expression levels in normal pancreas (n=18), CP (n=32), and PDAC tissue (n=33); (P<0.001 PDAC versus normal, P<0.001 PDAC versus CP).

Article Snippet: Tissue or cellular protein lysates (25 μg) and 75 μg of spleen tissue serving as a positive control, were separated by electrophoresis in 10-12% Bis-Tris gels (Invitrogen), transferred to nitrocellulose membrane and blocked with 5% non-fat dry milk in TTBS (20 mM Tris-HCl, 150 mM NaCl, and 0.1% Tween-20; 5% M-TTBS) for 1 h. The membranes were incubated overnight at 4 ̊C with goat anti-human CXCL16 (R&D Systems, Wiesbaden, Germany) or rabbit anti-human CXCR6 (ProSci, Poway, CA, USA), diluted 1:500 in 5% M-TTBS.

Techniques: Expressing, Quantitative RT-PCR

Journal: international Journal of Oncology

Article Title: Expression and potential function of the CXC chemokine CXCL16 in pancreatic ductal adenocarcinoma

doi: 10.3892/ijo_00000009

Figure Lengend Snippet: Figure 2. Expression of CXCL16 and CXCR6 protein in pancreatic cancer (PDAC) cells and pancreatic tissues. (A) CXCL16 and CXCR6 protein expression in pancreatic cancer cell lines. All tested cell lines displayed protein expression for CXCL16 (upper panel) and CXCR6 (lower panel). Spleen lysate served as a positive control. (B) CXCL16 protein expression in normal pancreas (n=4) and chronic pancreatitis (CP) (n=4) (lower panel), and pancreatic cancer (PDAC) tissue (n=8) (upper panel). Spleen lysate served as a positive control. In line with the results of the qRT-PCR, there is upregulated expression of CXCL16 protein in CP as well as in PDAC compared to normal pancreas. CXCR6 was not detectable in human tissue samples at the protein level using immunoblotting.

Article Snippet: Tissue or cellular protein lysates (25 μg) and 75 μg of spleen tissue serving as a positive control, were separated by electrophoresis in 10-12% Bis-Tris gels (Invitrogen), transferred to nitrocellulose membrane and blocked with 5% non-fat dry milk in TTBS (20 mM Tris-HCl, 150 mM NaCl, and 0.1% Tween-20; 5% M-TTBS) for 1 h. The membranes were incubated overnight at 4 ̊C with goat anti-human CXCL16 (R&D Systems, Wiesbaden, Germany) or rabbit anti-human CXCR6 (ProSci, Poway, CA, USA), diluted 1:500 in 5% M-TTBS.

Techniques: Expressing, Positive Control, Quantitative RT-PCR, Western Blot

Journal: international Journal of Oncology

Article Title: Expression and potential function of the CXC chemokine CXCL16 in pancreatic ductal adenocarcinoma

doi: 10.3892/ijo_00000009

Figure Lengend Snippet: Figure 3. Expression and localization of CXCL16 and CXCR6 in pancreatic tissues. Immunohistochemistry was performed as described in Materials and methods. CXCL16 immunoreaction in normal pancreas (A), chronic pancreatitis (CP) (B), and pancreatic cancer (PDAC) (C). CXCR6 immunoreaction in normal pancreas (D), CP (E), PDAC (F). Note the negative immunoreactivity in consecutive negative control tissue sections (inserts). The given slides reveal representative examples of the immunohistochemistry of CXCL16 and CXCR6.

Article Snippet: Tissue or cellular protein lysates (25 μg) and 75 μg of spleen tissue serving as a positive control, were separated by electrophoresis in 10-12% Bis-Tris gels (Invitrogen), transferred to nitrocellulose membrane and blocked with 5% non-fat dry milk in TTBS (20 mM Tris-HCl, 150 mM NaCl, and 0.1% Tween-20; 5% M-TTBS) for 1 h. The membranes were incubated overnight at 4 ̊C with goat anti-human CXCL16 (R&D Systems, Wiesbaden, Germany) or rabbit anti-human CXCR6 (ProSci, Poway, CA, USA), diluted 1:500 in 5% M-TTBS.

Techniques: Expressing, Immunohistochemistry, Negative Control

Journal: Frontiers in Neuroscience

Article Title: Teneurin-2 and related proteins in reactive astrocytes after status epilepticus induction in adult rats

doi: 10.3389/fnins.2025.1670634

Figure Lengend Snippet: Temporal immunoreactivity density percentage (pixels/area) analysis to ADGRL1-LI in the primary somatosensory area (A) and CA3 region of the hippocampus (B) in all experimental groups. Note a significant increase, principally in EG5 and EG14 compared to other groups. Mean (± SEM) values from each experimental group were submitted to one-way ANOVA and Tukey’s post hoc test, considering p < 0.05 as significant. In panel (A) , a, EG2 vs. EG14 ( p < 0.001); b, EG5 vs. NAG, PCG and EG65 ( p < 0.0001); c, EG5 vs. all other groups ( p < 0.0001); d, EG14 vs. NAG, PCG and EG65 ( p < 0.0001); e, EG14 vs. EG35 ( p < 0.01); f, EG35 vs. NAG and PCG ( p < 0.0001); g, EG65 vs. NAG and PCG ( p < 0.0001). In panel (B) , a, EG2 vs. NAG and PCG ( p < 0.0001); b, EG5 vs. NAG, PCG, EG2, EG35 and EG65 ( p < 0.0001); c, EG14 vs. NAG, PCG and EG60 ( p < 0.0001); d, EG14 vs. EG2 ( p < 0.05); e, EG14 vs. EG35 ( p < 0.01); f, EG14 vs. EG65 ( p < 0.001); g, EG35 vs. NAG, PCG ( p < 0.001); h, EG65 vs. NAG ( p < 0.01); EG65 vs. NAG ( p < 0.001). ADGRL1, adhesion G protein-coupled receptor L1; CA3, cornu mmonis 3 of the hippocampus; PCG, pharmacological control group; EG2, epilepsy group 2 days; EG5, epilepsy group 5 days; EG14, epilepsy group 14 days; EG35, epilepsy group 35 days; EG65, epilepsy group 65 days, NAG, naïve group.

Article Snippet: The sections were incubated in a solution of primary polyclonal antibody anti-Ten-2 raised in sheep (1:1000, CAH00223061, AF4578, R&D Systems, MN, USA), ADGRL1 raised in rabbit (1:1000, Lot ALR021AN0102, ALR-021, Alomone Labs, Jerusalem, Israel) and GFAP raised in chicken (1:500, Lot # 2987471, AB5541, EMD Millipore Corporation, CA, USA), diluted in PBS-TX and 2% normal donkey serum for 48 h at 4 °C with slow agitation.

Techniques: Control

Journal: Frontiers in Neuroscience

Article Title: Teneurin-2 and related proteins in reactive astrocytes after status epilepticus induction in adult rats

doi: 10.3389/fnins.2025.1670634

Figure Lengend Snippet: Indirect immunoperoxidase to ADGRL1-LI for PCG and all EG groups from frontal histologic sections of the adult rat hippocampus analyzed under light microscopy (A–F) . In panel (A) , note the subdivisions of the hippocampus (CA1, CA2, CA3 and dentate gyrus). In panel (E’) observe ADGRL1-LI reactive astrocytes in all layers of CA1 from EG35 and E” shows these cells distributed in stratum radiatum and stratum-lacunosum-moleculare. CA1, cornu Ammonis 1 of the hippocampus; CA2, cornu Ammonis 2 of the hippocampus; CA3, cornu Ammonis 3 of the hippocampus; D, dorsal; DG, dentate gyrus; L, lateral; M, medial; V, ventral.

Article Snippet: The sections were incubated in a solution of primary polyclonal antibody anti-Ten-2 raised in sheep (1:1000, CAH00223061, AF4578, R&D Systems, MN, USA), ADGRL1 raised in rabbit (1:1000, Lot ALR021AN0102, ALR-021, Alomone Labs, Jerusalem, Israel) and GFAP raised in chicken (1:500, Lot # 2987471, AB5541, EMD Millipore Corporation, CA, USA), diluted in PBS-TX and 2% normal donkey serum for 48 h at 4 °C with slow agitation.

Techniques: Light Microscopy

Journal: Frontiers in Neuroscience

Article Title: Teneurin-2 and related proteins in reactive astrocytes after status epilepticus induction in adult rats

doi: 10.3389/fnins.2025.1670634

Figure Lengend Snippet: Triple immunofluorescence of the hippocampus from EG5 epileptic group. Low magnification (A–E) and high magnification (F–J) of reactive astrocytes, exhibiting GFAP, Ten-2-LI, ADGRL1-LI and DAPI labeling under confocal microscopy. In panel (K) , a 3D reconstruction of a reactive astrocyte with triple immunolabeling is shown, identified by the arrows. ADGRL1, adhesion G protein-coupled receptor L1; CA2, cornu Ammonis 2 of the hippocampus; D, dorsal; GFAP, glial fibrillary acidic protein; L, lateral; M, medial; SO , stratum oriens ; SP, stratum pyramidale ; SR, stratum radiatum ; Ten-2, teneurin-2; V, ventral.

Article Snippet: The sections were incubated in a solution of primary polyclonal antibody anti-Ten-2 raised in sheep (1:1000, CAH00223061, AF4578, R&D Systems, MN, USA), ADGRL1 raised in rabbit (1:1000, Lot ALR021AN0102, ALR-021, Alomone Labs, Jerusalem, Israel) and GFAP raised in chicken (1:500, Lot # 2987471, AB5541, EMD Millipore Corporation, CA, USA), diluted in PBS-TX and 2% normal donkey serum for 48 h at 4 °C with slow agitation.

Techniques: Immunofluorescence, Labeling, Confocal Microscopy, Immunolabeling

Journal: Frontiers in Neuroscience

Article Title: Teneurin-2 and related proteins in reactive astrocytes after status epilepticus induction in adult rats

doi: 10.3389/fnins.2025.1670634

Figure Lengend Snippet: Temporal quantitative genic expression analysis to Ten-2 (A,B) , TCAP-2 (C,D) , and ADGRL1 (E,F) from primary somatosensory area and hippocampus samples ( n = 3, four animals per experimental group). Note the increase of gene expression to Ten-2 (A,B) , TCAP-2 (C,D) , and ADGRL1 (E,F) especially in the EG2 and EG5 groups. Mean (± SEM) values from each experimental group normalized in relation to GADPH and expressed in relation to NeuN (RBFOX3) submitted to one-way ANOVA and Tukey’s post hoc test, considering p < 0.05 as significant. In panel (A) , a EG5 vs. PCG and EG35 ( p < 0.05); b EG5 vs. NAG and EG14 ( p < 0.01). In panel (B) , a EG2 vs. NAG and PCG ( p < 0.01); b EG5 vs. NAG and PCG ( p < 0.01). In panel (C) , a EG5 vs. EG14 ( p < 0.05); b EG65 vs. EG14 ( p < 0.01). In panel (D) , a EG2 vs. EG65 (p < 0.05); b EG2 vs. NAG ( p < 0.001); c EG2 vs. PCG ( p < 0.01); d EG5 vs. EG65 ( p < 0.05); e EG5 vs. NAG and PCG ( p < 0.001); f EG14 vs. NAG and PCG ( p < 0.05). In panel (E) , a EG5 vs. NAG and PCG ( p < 0.05). In panel (F) , a EG2 vs. EG14 ( p < 0.05); b EG2 vs. NAG and PCG ( p < 0.0001); c EG2 vs. EG35 and EG6 ( p < 0.001); d EG5 vs. NAG and PCG ( p < 0.05); e EG5 vs. NAG and PCG ( p < 0.001). ADGRL1, adhesion G protein-coupled receptors L1; PCG, control group; EG2, epilepsy group 2 days; EG5, epilepsy group 5 days; EG14, epilepsy group 14 days; EG35, epilepsy group 35 days; EG65, epilepsy group 65 days; GHPDH, glyceraldehyde 3-phosphate dehydrogenase;; NeuN, neuron specific nuclear protein; RBFOX3, RNA binding Fox-1 homolog 3; TCAP-2, teneurin C-terminal-associated peptide 2; Ten-2, teneurin-2.

Article Snippet: The sections were incubated in a solution of primary polyclonal antibody anti-Ten-2 raised in sheep (1:1000, CAH00223061, AF4578, R&D Systems, MN, USA), ADGRL1 raised in rabbit (1:1000, Lot ALR021AN0102, ALR-021, Alomone Labs, Jerusalem, Israel) and GFAP raised in chicken (1:500, Lot # 2987471, AB5541, EMD Millipore Corporation, CA, USA), diluted in PBS-TX and 2% normal donkey serum for 48 h at 4 °C with slow agitation.

Techniques: Expressing, Gene Expression, Control, RNA Binding Assay