Journal: BMC Cancer

Article Title: CDC6, a key replication licensing factor, is overexpressed and confers poor prognosis in diffuse large B-cell lymphoma

doi: 10.1186/s12885-023-11186-6

Figure Lengend Snippet: CDC6 regulates downstream signalings related to cell cycle and apoptosis. A and B, SUHDL4 cells ( A ) and OCI-LY7 ( B ) were transfected with pLenO vector (LV-NC), pLenO-CDC6 (LV-CDC6), pLenR-GPH vector (LV-shCtrl) and pLenR-shCDC6 (LV-shCDC6) for 72 h, respectively, and the levels of CDC6, caspase-3, Bcl-2, Bax and GAPDH were analyzed by Western blot analysis. C and D, the experiments were performed as described in A and B , the cell lysates were collected and analyzed by western blot with antibodies against INK4, E-Cadherin, ATR, and GAPDH, respectively

Article Snippet: The primary antibodies against CDC6 (ab109315), caspase-3 (ab197202), bcl-2 (ab182858), Bax (ab182734), E-cadherin (ab40772), and ATR (ab13798) were purchased from abcam, UK; anti-INK4 (Cst 74,560) was from Cell Signaling Technology, US.

Techniques: Transfection, Plasmid Preparation, Western Blot

Journal: Neoplasia (New York, N.Y.)

Article Title: Multi-omics analysis unveils tumor heterogeneity and immunotherapy predictive model in breast cancer for precision medicine and early detection

doi: 10.1016/j.neo.2025.101260

Figure Lengend Snippet: Expression and functional analysis of PIS in breast cancer. (A) Relative mRNA expression of PIS in breast cancer tissues and matched adjacent normal tissues. (B) PIS mRNA expression levels in breast cancer cell lines (MDA-MB-361, T-47D, MCF7, MD-AMB-436, BT-483) related to MCF-10A. (C) qRT-PCR validation of PIS knockdown efficiency in MCF7 and MDA-MB-436 cells following siRNA transfection. (D–E) CCK-8 assays showing cell proliferation over 4 days in control and PIS knockdown groups (n = 5 per group). (F–G) Flow cytometry analysis of apoptosis in control and PIS-silenced MDA-MB-436 cells. (H–I) Transwell migration and invasion assays in MDA-MB-436 cells following PIS knockdown. (J–K) Western blot analysis of cleaved caspase-3, Bcl-2, E-cadherin, and Vimentin expression in control and PIS knockdown MDA-MB-436 cells.

Article Snippet: After blocking with 5% non-fat milk for 1 hour at room temperature, membranes were incubated overnight at 4°C with the following primary antibodies: cleaved Caspase-3 (1:1000, #9664, CST), E-Cadherin (1:2000, #3195, CST), Bcl-2 (1:1000, #15071, CST), Vimentin (1:1000, #5741, CST), and β-actin (1:5000, #4970, CST).

Techniques: Expressing, Functional Assay, Quantitative RT-PCR, Biomarker Discovery, Knockdown, Transfection, CCK-8 Assay, Control, Flow Cytometry, Migration, Western Blot

Journal: Oncology reports

Article Title: Long non‑coding RNA PVT1 promotes epithelial‑mesenchymal transition via the TGF‑β/Smad pathway in pancreatic cancer cells.

doi: 10.3892/or.2018.6462

Figure Lengend Snippet: Figure 6. PVT1 enhances EMT via TGF‑β/Smad signaling. (A and B) Immunoblotting of Smad2/3, p‑Smad2/3, Smad4, TGF‑β1 in PaTu8988 and BxPc‑3 cells treated with siPVT1‑NC, siPVT1‑1, siPVT1‑2. (C) Immunoblotting of Smad2/3, p‑Smad2/3, TGF‑β1 in SW1990 cells transfected with pCD513B‑1‑PVT1 or pCD513B‑1. β‑tubulin was used as a loading control. *P<0.05. Data are expressed as the mean ± SD from three independent experiments. (D and E) TGF‑β1 knockdown downregulated p‑Smad2/3 after BxPc‑3 and SW1990 cells were transfected with pCD513B‑1‑PVT1.

Article Snippet: The antibodies used were: rabbit anti-TGF-β (cat. no. 3712; Cell Signaling Technology, Inc., Danvers, MA, USA), rabbit anti‐MMP2 (cat. no. YT2798) and rabbit anti‐MMP9 (cat. no. YT1892; both from ImmunoWay Biotechnology Co., Plano, TX, USA), an EMT antibody sampler kit (cat. no. 9782), a Smad2/3 antibody sampler kit (cat. no. 12747), mouse anti-β-Tubulin (cat. no. 4466), and rabbit anti-β‐actin (cat. no. 4970; all from Cell Signaling Technology, Inc.) and all above antibodies diluted with 1:1,000. β-actin and β-tubulin were used as internal controls.

Techniques: Western Blot, Transfection, Control, Knockdown