primary antibodies against ki67  (Cell Signaling Technology Inc)

Journal: iScience

Article Title: HHEX-PRKAR2B axis-mediated PKA activation drives glucose metabolism-dependent progression of pancreatic ductal adenocarcinoma

doi: 10.1016/j.isci.2026.114691

Figure Lengend Snippet: In vivo demonstration of glucose-activated PKA promoting glycolysis-dependent progression A. Immunoblot analysis of PRKAR2B in KPC cells with or without PRKAR2B knockout. (B–D) Representative images of colony formation by KPC cells, with or without PRKAR2B knockout, cultured in normal or low-glucose medium (B). Colony size (C) and number of colonies (D) were graphed ( n = 3 independent repeats). Data were expressed as mean ± SD. Scale bars, 50 μm ∗, p < 0.05, ∗∗, p < 0.01, NS, no significance. (E) Representative H&E-stained pancreatic sections from mice with tamoxifen-inducible, pancreas-specific KRAS expression, harvested 3 weeks after tamoxifen or vehicle injection. Scale bars, 1 mm (F–H) PKA activity (F), pyruvic acid level (G), and lactate level (H) in pancreatic tissues from mice, harvested 3 weeks after tamoxifen or vehicle injection, assessing the effect of tamoxifen-inducible, pancreas-specific KRAS expression ( n = 4 independent samples). Data were expressed as mean ± SD. ∗, p < 0.05; ∗∗, p < 0.01. (I) Colorimetric analysis of PKA activity in wild-type and diabetic mice ( n = 4 independent samples). Data were expressed as mean ± SD. ∗∗, p < 0.01. (J) Representative immunoblot of CREB, pCREB, and KLF5 in wild-type and diabetic mice ( n = 4 independent samples; see F for all replicates). (K) mRNA expression level of HK2 in wild-type and diabetic mice ( n = 4 independent samples). Data were expressed as mean ± SD. ∗∗, p < 0.01. (L) Schematic diagram of the experimental design in mice. Pan02 cells with or without PRKAR2B knockout was injected into wild type or diabetic mouse through the tail vein ( n = 4 independent samples). (M) Changes in mouse body weight following tail vein injection of PRKAR2B -knockout or wild-type Pan02 cells ( n = 4 independent samples). Data were expressed as mean ± SD. ∗∗, p < 0.01. (N) Colorimetric analysis of PKA activity in tumor allografts from wild-type and diabetic mice with or without PRKAR2B knockout ( n = 4 independent samples). Data were expressed as mean ± SD. ∗, p < 0.05, ∗∗, p < 0.01. (O) Representative image of Ki67 IHC staining in Pan02-control and Pan02- PRKAR2B -sg allografts from wild-type or diabetic mice ( n = 4 independent samples). Scale bars, 150 μm (P) Representative H&E-stained lung sections at the experimental endpoint. Scale bars, 3 mm (Q–R) Quantification of metastatic lesion number (Q) and total metastatic burden (R) at the experimental endpoint across the indicated treatment groups ( n = 4 independent samples). Data were expressed as mean ± SD. ∗, p < 0.05; NS, no significance.

Article Snippet: Primary antibody against Ki67 (1:500 dilution, cat. 12202, Cell Signaling Technology, MA, USA) was prepared in SignalStain Antibody Diluent (cat. 8112, Cell Signaling Technology) and applied to the sections overnight at 4°C.

Techniques: In Vivo, Western Blot, Knock-Out, Cell Culture, Staining, Expressing, Injection, Activity Assay, Immunohistochemistry, Control

Journal: BMC Cancer

Article Title: MiR-218-5p inhibits glioma progression by targeting CKLF-like MARVEL transmembrane domain-containing 3

doi: 10.1186/s12885-026-15544-y

Figure Lengend Snippet: Cell counting kit-8 ( A ) and EdU ( B – C ) assays were used to analyze the proliferation of CMTM3-knockdown, si-NC-transfected, and control U251 and U87 cells. Data are expressed as the mean ± SD of three independent experiments (*, P < 0.05; **, P < 0.01; ns, not significant) (scale bar = 100 μm). D Quantitative analysis of bioluminescence images of the CMTM3 shRNA and negative control shRNA (sh-NC) treatment groups ( n = 6). Statistical analysis revealed notable differences between the treatment groups and the control group. E The survival rate of nude mice injected with U87 cells transfected with lentiviruses encoding either sh-NC or sh-CMTM3 was assessed using Kaplan–Meier curves. F The bioluminescence activity of nude mice injected with U87 cells was assessed on days 10 and 21 after implantation. G Immunohistochemical analysis showed reduced CMTM3 and Ki67 expressions in brain slices with tumor tissue of nude mice injected with sh-CMTM3-transfected U87 cells (*, P < 0.05; **, P < 0.01) (scale bar = 20 μm)

Article Snippet: After antigen retrieval, the sections were blocked with 5% goat serum and 3% hydrogen peroxide and incubated with specific primary antibodies against Ki67 (1:100; ProteinTech Group, Rosemont, IL, USA) and CMTM3 (1:100; Thermo Fisher Scientific).

Techniques: Cell Counting, Knockdown, Transfection, Control, shRNA, Negative Control, Injection, Activity Assay, Immunohistochemical staining