antigens cyclin d1  (Vector Laboratories)

Journal: Autoimmunity

Article Title: LncRNA NONHSAT042241 inhibits rheumatoid synovial proliferation, inflammation and aggression via inactivating WNT/β-catenin signaling pathway.

doi: 10.1080/08916934.2024.2387076

Figure Lengend Snippet: Figure 4. LLDT-8 Inhibited activation of wnt/β-catenin pathway in RA-FLS via NONHSAT042241. (A) Protein expression of wnt/β-catenin pathway key members, including p-GSK-3β, and β-catenin in various cell groups. (B) Protein expression of PCNA and CyclinD1 in various cell groups. Compared with control group, **p < 0.01; compared with OE-lnc 2241 group.

Article Snippet: The primary antibodies included an anti-proliferating cell nuclear antigen (PCNA) antibody (Cell Signaling Technology), an anti-cyclin D1 antibody (Cell Signaling Technology), an anti-Bax antibody (Cell Signaling Technology), an anti-Cleaved caspase-3 antibody (Cell Signaling Technology), an anti-β-Catenin antibody (Abcam Cell Signaling Technology), an anti-GSK -3β antibody (Cell Signaling Technology), anti-p-GSK3βantibodies (Cell Signaling Technology), and a GAPDH and β-actin antibody (Cell Signaling Technology).

Techniques: Activation Assay, Expressing, Control

Journal: Cells

Article Title: CD133 Stimulates Cell Proliferation via the Upregulation of Amphiregulin in Melanoma

doi: 10.3390/cells13090777

Figure Lengend Snippet: Two melanoma cell lines BAKP and POT, exhibit higher levels of phosphorylation and activation of EGFR and members of the MAPK pathway (p-MEK, p-ERK, p-BAD compared to total MEK, ERK, BAD) ( A ) as well as the cell proliferation marker PCNA and cyclin D1 ( B ) in Dox-induced vs. uninduced cells. ( A ) Cell lysates of BAKP and POT cells incubated with or without 1 µg/mL Dox for 24 h were subjected to immunoblot analysis with antibodies to pEGFR. Immunoblots were stripped of antibodies and re-probed with antibodies to p-MEK, MEK, p-ERK, ERK, p-BAD, and β-actin for loading control. ( B ) Cell lysates of BAKP and POT cells incubated with or without 1 µg/mL Dox for 24 h were subjected to immunoblot analysis with antibodies to the proliferation marker PCNA, then β-actin for loading control. ( C ) BAKP cells incubated with or without 1 µg/mL Dox for 24 h were subjected to Western blot analysis with antibodies to CD133, then stripped of antibodies and re-probed with antibodies to TOP2 and β-actin, as loading control. Immunoblots loaded with the same lysates were likewise probed with antibodies to Cyclin D1, followed by β-actin. Densitometric analysis is shown in immunoblots, comparing intensities of protein bands relative to that of control −Dox cells, after normalizing to β-actin.

Article Snippet: Membranes were then incubated with antibodies to CD133 (Miltenyi Biotec, Auburn, CA, USA), AREG (Santa Cruz Biotech, Dallas, TX, USA), p-EGFR, p-MEK, p-BAD (BioLegend, San Diego, CA, USA), p-ERK, total MEK, ERK, BAD, proliferating cell nuclear antigen (PCNA), cyclin D1, or β-actin (ProteinTech, Rosemont, IL, USA) as loading control.

Techniques: Phospho-proteomics, Activation Assay, Marker, Incubation, Western Blot, Control

Journal: Cells

Article Title: CD133 Stimulates Cell Proliferation via the Upregulation of Amphiregulin in Melanoma

doi: 10.3390/cells13090777

Figure Lengend Snippet: CD133 stimulates cell proliferation and promotes melanoma progression via a novel CD133-AREG-EGFR-MAPK activation pathway in CD133-positive melanoma-initiating stem cells (MICs), summarized as follows: (1) CD133 upregulates AREG, (2) AREG is cleaved to its ligand form by metalloproteinase ADAMS, (3) AREG ligand binds to and activates EGFR, (4) EGFR activates the MAPK pathway (by phosphorylating MEK, which phosphorylates ERK), (5) ERK phosphorylates and inactivates apoptotic protein BAD, leading to increased cell survival, (6) activation of the MEK/ERK pathway upregulates cyclin D1, (7) resulting in activation of E2F1, 8), which in turn binds to and upregulates the S-phase gene promoters, including those of PCNA.

Article Snippet: Membranes were then incubated with antibodies to CD133 (Miltenyi Biotec, Auburn, CA, USA), AREG (Santa Cruz Biotech, Dallas, TX, USA), p-EGFR, p-MEK, p-BAD (BioLegend, San Diego, CA, USA), p-ERK, total MEK, ERK, BAD, proliferating cell nuclear antigen (PCNA), cyclin D1, or β-actin (ProteinTech, Rosemont, IL, USA) as loading control.

Techniques: Activation Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: The effects of patchouli alcohol and combination with cisplatin on proliferation, apoptosis and migration in B16F10 melanoma cells

doi: 10.1111/jcmm.17745

Figure Lengend Snippet: The expression levels of cell cycle and apoptosis relative proteins regulated by PA treatment. Cells were incubated with 89.9 μM of PA for 0, 6, 12, 24 and 48 h, and the expression level of proteins associated with cell cycle and apoptosis was detected using western blotting. PA treatment dramatically attenuated the expressions of PCNA, cyclin D1 and CDK4, while activating caspase‐8, caspase‐9 and caspase‐3. Data are represented as means ± SD of at least three independent experiments. * P < 0.05, versus control with a significant increase. # P < 0.05, versus control with a significant decrease. C‐Cas, cleaved‐caspase; PCNA, proliferating cell nuclear antigen; Pro‐Cas, pro‐caspase.

Article Snippet: For western blotting and IHC staining experiments, primary antibodies against proliferating cell nuclear antigen (PCNA), cyclin D1, cyclin‐dependent kinase 4 (CDK4), caspase‐8, caspase‐9, caspase‐3, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)‐2 and MMP‐9 were purchased from Santa Cruz Biotechnology; Smad2/3, p‐Smad2/3 were purchased from Cell Signalling Technology; E‐cadherin, vimentin and β‐actin were purchased from iReal Biotechnology Co., Ltd.

Techniques: Expressing, Incubation, Western Blot, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: The effects of patchouli alcohol and combination with cisplatin on proliferation, apoptosis and migration in B16F10 melanoma cells

doi: 10.1111/jcmm.17745

Figure Lengend Snippet: PA regulated markers expression of proliferation, apoptosis, angiogenesis and metastasis in vivo. The mice (vehicle and 150 mg/kg PA groups) were sacrificed when the tumour volume exceeded 1500 mm 3 , and then the tumour was fixed, sliced and stained with H&E (scale bar: 100 μm), IHC (scale bar: 50 μm) and TUNEL (scale bar: 50 μm) for histological examination. Image of H&E staining showed the morphology of cell death and nucleolysis (arrow). The expression of caspase‐3, PCNA, VEGF, MMP‐2 and MMP‐9 were examined using immunohistochemical analysis and scored using the Quickscore method. PA‐induced cell apoptosis was determined using tissue TUNEL stain. All data are shown as mean values ± SEM. * P < 0.05, versus vehicle group. H&E, haematoxylin and eosin; IHC, immunohistochemistry; MMP, matrix metalloproteinases; PCNA, proliferating cell nuclear antigen; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labelling; VEGF, vascular endothelial growth factor.

Article Snippet: For western blotting and IHC staining experiments, primary antibodies against proliferating cell nuclear antigen (PCNA), cyclin D1, cyclin‐dependent kinase 4 (CDK4), caspase‐8, caspase‐9, caspase‐3, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)‐2 and MMP‐9 were purchased from Santa Cruz Biotechnology; Smad2/3, p‐Smad2/3 were purchased from Cell Signalling Technology; E‐cadherin, vimentin and β‐actin were purchased from iReal Biotechnology Co., Ltd.

Techniques: Expressing, In Vivo, Staining, TUNEL Assay, Immunohistochemical staining, Immunohistochemistry

Journal: Antioxidants (Basel, Switzerland)

Article Title: Antrodin C Isolated from Antrodia Cinnamomea Induced Apoptosis through ROS/AKT/ERK/P38 Signaling Pathway and Epigenetic Histone Acetylation of TNFα in Colorectal Cancer Cells.

doi: 10.3390/antiox12030764

Figure Lengend Snippet: Figure 5. Antrodin C (ADC) varies proliferative and apoptotic protein expression in CRC cells in an in vivo xenograft mouse model. Hematoxylin and eosin staining of the tumors of the nude mice was performed (1st row, upper panel). Protein levels of PCNA (2nd row, upper panel), cyclin D1 (3rd row, upper panel), cyclin E (5th row, upper panel), MMP-9 (6th row, upper panel), and TNFα (7th row, upper panel) were detected by using immunohistochemical analysis. Right panel: By calculating the average of integrated optical density (AIOD), quantitative assay of PCNA, cyclin D1, cyclin E, MMP-9, and TNFα proteins was evaluated. Multiple tumor fields and their positively stained area were evaluated and then evaluated by randomly selecting three observational fields of each section (n = 6/group). The data are expressed as mean ± SD. * p < 0.05, magnification × 200.

Article Snippet: In addition to immunohistochemical analysis, every subcutaneous tumor specimen was first blocked, then incubated with monoclonal antiproliferating cell nuclear antigen (PCNA), cyclin D1, cyclin E, matrix metalloproteinase (MMP)-9, and TNFα antibodies (Santa Cruz, CA, USA) overnight.

Techniques: Expressing, In Vivo, Staining, Immunohistochemical staining