Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Detection of β-catenin, E- and N-cadherins and the cellular localization of the proteins in chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal liver (control)

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Control

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Immunohistochemical ( A – C ) and hybridocytochemical localization of β-catenin ( D ) in liver. Membranous/ cytoplasmic localization of β-catenin in liver with chronic hepatitis C ( A ), membranous localization of β-catenin in hepatocellular carcinoma ( B ) and in control liver ( C ); mRNA for β-catenin in cytoplasm and cell nuclei in HCC fragment ( D ). Immunocytochemical detection of E-cadherin ( E – F ) and N-cadherin ( H ) in liver. Predominantly membranous localization of E-cadherin in liver with chronic hepatitis C ( E ), hepatocellular carcinoma ( F ) and control liver ( G ). Membrano-cytoplasmic localization of N-cadherin in fragment of hepatocellular carcinoma ( H ). Immunohistochemistry ( A – C , E – H ) technique and hybridization in situ method ( D ). Hematoxylin counterstained. Bar = 40 μm

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Immunohistochemical staining, Control, Immunohistochemistry, Hybridization, In Situ

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Comparison of quantitatively assessed β-catenin, E-cadherin and N-cadherin immunoexpression (% of the IHC reaction area in analysed area of liver parenchyma) in chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal liver (C)

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Comparison

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Comparative immunoexpression of β-catenin, E-cadherin and N-cadherin in liver with chronic hepatitis C (CH-C), hepatocellular carcinoma (HCC) and normal organ (control) ***p (level of significance) value < 0.001, *p < 0.05.

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Control

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Tissue expression of β-catenin, E-cadherin and N-cadherin (mean % of IHC reaction area ± SD) as related to grading and staging in chronic hepatitis C (CH-C) group

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Expressing

Journal: Archives of Medical Science : AMS

Article Title: Tissue expression of β-catenin and E- and N-cadherins in chronic hepatitis C and hepatocellular carcinoma

doi: 10.5114/aoms.2017.65272

Figure Lengend Snippet: Values of Spearman’s rank coefficient between expression of β-catenin, E-cadherin, N-cadherin (% IHC reaction area per area of hepatic parenchyma) and clinicopathological data in CH-C group

Article Snippet: Mouse anti-human monoclonal antibodies (mAbs) were employed, directed against human beta-catenin (Clone 196618; Code M 7052) (in dilution 1 : 50) (R&D Systems, UK), against E-cadherin (clone NCH-38) (in dilution 1 : 100) (DakoCytomation, Gdynia, PL) and against N-cadherin (in dilution 1 : 50) (clone IAR06) (Novocastra Labs. Ltd, Newcastle upon Tyne, UK).

Techniques: Expressing, Infection

Journal: Cell death and differentiation

Article Title: CDK15 promotes colorectal cancer progression via phosphorylating PAK4 and regulating β-catenin/ MEK-ERK signaling pathway.

doi: 10.1038/s41418-021-00828-6

Figure Lengend Snippet: Fig. 5 PAK4 mediates the oncogenic effect of CDK15 in colorectal cancer. A p-β-catenin(Ser675), c-Myc, p-MEK1/2 (Ser217/221), and p-ERK1/ 2 (Thr202/Tyr204) were detected by western blot after CDK15 knockdown in CCD18-co, SW480 and HCT116 cells. B Cells with PAK4 silencing and CDK15 overexpression were established. PAK4 and CDK15 expression was determined by western blot. C PAK4 knockdown reverses cell proliferation induced by CDK15 in CCD18-co and HCT116 cells. MTT assay was used to detect cell proliferation. D Anchorage-independent growth in CCD18-co and HCT116 cells with PAK4 silencing and CDK15 overexpression. Left panels: representative images (Scale bar: 200 μm). Right panels: Colonies were counted using Image J-Plus (Scale bar: 200 μm) and data represented statistical analysis of colony number ratio. E Anchorage-independent growth in CCD18-co and HCT116 cells treated with PAK4 inhibitor (PF-3758309). Left panel: representative images of colonies (Scale bar: 200 μm). Right panel: statistical analysis of the colony ratio. F Western blot to validate β-catenin and MEK/ERK signaling pathway in HCT116 cells with indicated treatment. Data were presented as mean values ± SD from triplicate experiments. Statistical differences were evaluated using Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The following antibodies were used in our study: anti-CDK15 (Cat#PA5-28595, Invitrogen), anti-CDK15 (Cat#TA811934, ORIGENE), anti-PAK4 (Cat#sc-390507, Santa Cruz), anti- phospho-β-catenin (Ser675) (Cat#4176, Cell Signaling Technology), anti-β-catenin (Cat # 8480, Cell Signaling Technology), antiphospho-ERK1/2 (Thr202/Tyr204)(Cat# 4370, Cell Signaling Technology), Anti-ERK1/2 (Cat#4695, Cell Signaling Technology), anti-phospho-MEK1/2 (Ser217/221)(Cat# 9154, Cell Signaling Technology), anti-MEK1/2 (Cat# 4694, Cell Signaling Technology), anti-c-Myc antibody (Cat#ab32072, Abcam), anti-Flag (Cat #F1804, Sigma), anti-HA (Cat #3724, Cell Signaling Technology), anti-His (Cat #ab137839, Abcam), anti-GAPDH (#HRP-60004, Proteintech), anti-β-actin (#HRP-60008, Proteintech).

Techniques: Western Blot, Knockdown, Over Expression, Expressing, MTT Assay

Journal: Cell death and differentiation

Article Title: CDK15 promotes colorectal cancer progression via phosphorylating PAK4 and regulating β-catenin/ MEK-ERK signaling pathway.

doi: 10.1038/s41418-021-00828-6

Figure Lengend Snippet: Fig. 7 Targeting PAK4 delays tumor growth in patient-derived xenografts. A Clinical information for HJG208 and HJG210 from patient’s cancer tissues. B, C 0.9% NaCl as vehicle, 5 mg/kg or 20 mg/kg PF-3758309 were intraperitoneally injected once per day for 20 days, and tumor volume was monitored every 2–5 days (n = 9–10 mice per group). D, E Tumor photographs. F, G Tumor weight and tumor growth inhibition (H, I) normalized to control group. J Levels of p-β-catenin (Ser675), c-Myc, p-MEK1/2 (Ser217/221), and p-ERK1/2 (Thr202/Tyr204) in harvested tumor tissues were assessed by immunohistochemistry. Representative photographs for each antibody in different groups are shown (100×; Scale bar: 50 μm). K Statistical analysis for immunohistochemistry staining. Statistical differences were evaluated using Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001. Error bars represent mean ± SD.

Article Snippet: The following antibodies were used in our study: anti-CDK15 (Cat#PA5-28595, Invitrogen), anti-CDK15 (Cat#TA811934, ORIGENE), anti-PAK4 (Cat#sc-390507, Santa Cruz), anti- phospho-β-catenin (Ser675) (Cat#4176, Cell Signaling Technology), anti-β-catenin (Cat # 8480, Cell Signaling Technology), antiphospho-ERK1/2 (Thr202/Tyr204)(Cat# 4370, Cell Signaling Technology), Anti-ERK1/2 (Cat#4695, Cell Signaling Technology), anti-phospho-MEK1/2 (Ser217/221)(Cat# 9154, Cell Signaling Technology), anti-MEK1/2 (Cat# 4694, Cell Signaling Technology), anti-c-Myc antibody (Cat#ab32072, Abcam), anti-Flag (Cat #F1804, Sigma), anti-HA (Cat #3724, Cell Signaling Technology), anti-His (Cat #ab137839, Abcam), anti-GAPDH (#HRP-60004, Proteintech), anti-β-actin (#HRP-60008, Proteintech).

Techniques: Derivative Assay, Injection, Inhibition, Control, Immunohistochemistry, Staining

Journal: Cell death and differentiation

Article Title: CDK15 promotes colorectal cancer progression via phosphorylating PAK4 and regulating β-catenin/ MEK-ERK signaling pathway.

doi: 10.1038/s41418-021-00828-6

Figure Lengend Snippet: Fig. 8 Lentivirus-mediated CDK15 silencing inhibits colorectal tumor growth in patient-derived xenografts. A Clinical information for HJG86 from patient’s cancer tissues. B Mice received the lentiviruses (shNT, shCDK15-3, shCDK15-7) via intratumoral injection every 3 days for a total of four times. Tumor volume was monitored every 2–5 days for four continuous weeks (n = 8 mice per group). C Tumors photographs. D Tumor weight measured at the end of the study. E Tumor growth inhibition normalized to control group. F Levels of CDK15, p-β-catenin (Ser675), c-Myc, p-MEK1/2(Ser217/221), and p-ERK1/2 (Thr202/Tyr204) in harvested tissues were assessed by immunohistochemistry. Representative photographs in different groups are shown (100×; Scale bar: 50 μm). G Statistical analysis for immunohistochemistry staining. H Schematic model for the findings of this work: Aberrant CDK15 in CRC binds PAK4 and phosphorylates PAK4 at the S291 site. Accumulated phosphorylation of S291 upregulates the β-catenin/c-Myc and MEK/ERK signals, which in turn contribute to CRC tumor growth. Statistical differences were evaluated using Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001. Error bars represent mean ± SD.

Article Snippet: The following antibodies were used in our study: anti-CDK15 (Cat#PA5-28595, Invitrogen), anti-CDK15 (Cat#TA811934, ORIGENE), anti-PAK4 (Cat#sc-390507, Santa Cruz), anti- phospho-β-catenin (Ser675) (Cat#4176, Cell Signaling Technology), anti-β-catenin (Cat # 8480, Cell Signaling Technology), antiphospho-ERK1/2 (Thr202/Tyr204)(Cat# 4370, Cell Signaling Technology), Anti-ERK1/2 (Cat#4695, Cell Signaling Technology), anti-phospho-MEK1/2 (Ser217/221)(Cat# 9154, Cell Signaling Technology), anti-MEK1/2 (Cat# 4694, Cell Signaling Technology), anti-c-Myc antibody (Cat#ab32072, Abcam), anti-Flag (Cat #F1804, Sigma), anti-HA (Cat #3724, Cell Signaling Technology), anti-His (Cat #ab137839, Abcam), anti-GAPDH (#HRP-60004, Proteintech), anti-β-actin (#HRP-60008, Proteintech).

Techniques: Derivative Assay, Injection, Inhibition, Control, Immunohistochemistry, Staining, Phospho-proteomics

Journal: Cell Death Discovery

Article Title: Monomethyl fumarate confers cardioprotection after myocardial infarction via HCAR2-dependent activation of PI3K/Akt signaling

doi: 10.1038/s41420-025-02927-6

Figure Lengend Snippet: A Immunofluorescence staining of transverse LV sections for HCAR2 (red) and β-catenin (green) with DAPI nuclear stain (blue), shown individually and merged. Sham hearts show strong HCAR2 expression throughout the myocardium. MI hearts have markedly diminished HCAR2 signal (especially in the infarct zone; yellow arrows indicate areas of lost HCAR2 in MI). MMF-treated MI hearts retain higher HCAR2 expression (red fluorescence is partially preserved in the LV wall). β-catenin staining (green) labels cell-cell junctions and was used as a counterstain; nuclei are blue. Scale bar = 2 mm (whole section). B High-magnification images of myocardium from the same groups stained for HCAR2 (red), β-catenin (green), and DAPI (blue). These panels highlight HCAR2 at the cellular level. In sham myocardium, HCAR2 is readily detected in cardiomyocytes (red); in MI myocardium, HCAR2 is nearly absent; with MMF, HCAR2 signal is visibly greater than MI alone. Scale bar = 50 µm. C Mean fluorescence intensity of HCAR2 immunostaining in myocardium (arbitrary units). MI significantly reduces HCAR2 intensity vs Sham (# p < 0.05, n = 4), while MMF treatment results in a higher HCAR2 signal compared to MI (* p < 0.05, n = 4). D Western blots of key proteins in LV tissue: phosphorylated Akt (p-Akt, Ser473), total Akt, HCAR2, and GAPDH. Each lane represents a different animal per group. MI causes a loss of HCAR2 protein and a major decrease in p-Akt levels relative to sham. MMF-treated hearts show increased p-Akt and higher HCAR2 protein compared to MI. E , F Quantification of Western blot data (band intensity ratios, Sham set to 1.0). E p-Akt/Akt ratio in each group. Akt phosphorylation is greatly diminished after MI (# p < 0.05 vs Sham, n = 4), but significantly elevated in the MMF group (* p < 0.05 vs MI, n = 4), indicating restored Akt activity. F HCAR2/GAPDH protein ratio. MI hearts exhibit dramatically lower HCAR2 levels than Sham (# p < 0.05, n = 4), whereas MI + MMF hearts have significantly higher HCAR2 expression than MI (* p < 0.05, n = 4). These data suggest that HCAR2 and downstream Akt signaling are suppressed by MI injury and that MMF prevents this suppression, potentially linking HCAR2/Akt to MMF’s protective effects. Symbols: # p < 0.05 vs Sham; * p < 0.05 vs MI. MMF monomethyl fumarate, MI myocardial infarction, DAPI 4’,6-diamidino-2-phenylindole, HCAR2 Hydroxycarboxylic Acid Receptor 2, GAPDH glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: Slides were then incubated overnight at 4 °C with primary antibodies against HCAR2 (rabbit polyclonal, 1:200, Invitrogen; # PA5-90579) and β-catenin (mouse monoclonal, 1:200, Cell Signaling Technology; #37447).

Techniques: Immunofluorescence, Staining, Expressing, Fluorescence, Immunostaining, Western Blot, Phospho-proteomics, Activity Assay