normal colon epithelial cell line ncm460  (ATCC)

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: The E3 Ubiquitin Ligase ARIH1 Facilitates Colorectal Cancer Progression by Promoting Oxidative Phosphorylation via the Mitochondrial Translocation of K63-Linked Ubiquitinated PHB1.

doi: 10.1002/advs.202501017

Figure Lengend Snippet: Figure 1. ARIH1 was highly-expressed in CRC tissues and cell lines and correlated with poor prognosis. A) The ARIH1 mRNA expression levels were quantified by qRT-PCR in 30 CRC tissues and matched adjacent normal tissues. B-E) The protein level of ARIH1 detected by IHC in the TMA of 80 CRC patients’ samples. B) Comprehensive tissue microarray schema of ARIH1 expression, C) representative images of ARIH1 expression, D) the relative statistical analysis of H-score, and E) the Kaplan-Meier survival analysis of patients with high (n = 35) and low (n = 45) ARIH1 expression. F) The protein level of ARIH1 was detected by WB in 12 paired CRC tumor and adjacent tissues. G) The mRNA and protein levels of ARIH1 in CRC cell lines and NCM460 cells, evaluated by qRT-PCR and WB respectively. Data was shown as mean±SD of three independent experiments, *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Cell Culture: The human normal colonic epithelial cell line NCM460, HEK293T, along with CRC cell lines, including HCT116, DLD-1, SW620, SW480, LOVO, and RKO, were purchased from Servicebio Technology (Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Microarray

Journal: Neural Regeneration Research

Article Title: Elucidation of the mechanism by which manganese-ferric Prussian blue nanozymes alleviate ischemic stroke damage in a mouse model

doi: 10.4103/nrr.nrr-d-24-00837

Figure Lengend Snippet: Figure 6 ｜Therapeutic mechanisms of MnPB nanozymes and SCFAs in MCAO model. (A) Illustration of the concentration-dependent cellular uptake of MnPB nanozymes, highlighting the biocompatibility and internalization efficiency in NCM460 cells. Scale bar: 50 μm. (B) Assessment of the toxic effects of MnPB nanozymes on NCM460 cells, demonstrating safety within a concentration range of 0–50 μg/mL (n = 5). (C) Assessment of the toxic effects of MnPB nanozymes on BV-2 cells, demonstrating safety within a concentration range of 0–50 μg/mL (n = 5) (D) Examination of the toxic impact of H2O2 on BV-2 cells, establishing a baseline for oxidative stress-induced injury (n = 5). (E) Evaluation of the protective effects of different concentrations of MnPB nanozymes and SCFAs on cell survival in an in vitro oxidative stress injury model induced by H2O2 (n = 5). (F–H) Determination of the cytotoxicity of different concentrations of sodium acetate (NaA), sodium propionate (NaP), and sodium butyrate (NaB) on BV-2 cells after a 12-hour treatment period using the CCK-8 kit (n = 5). (I) Assessment of the combined toxicity of NaA, NaP, NaB, and SCFAs mixtures on BV-2 cells after a 12-hour exposure, confirming the safety of the selected concentration ratios (n = 5). (J) Utilization of the DCFH-DA assay to measure the reactive oxygen species (ROS) scavenging effect of varying concentrations of SCFAs. (K) Quantitative analysis of ROS spectral intensity, providing insights into the antioxidant capacity of SCFAs (n = 3). (L) Flow cytometry results depicting the apoptotic effects in BV-2 cells under oxidative stress. (M) Statistical analysis of apoptosis rates, revealing the protective role of SCFAs against cell death (n = 3). (N) Analysis of LPS-induced inflammation in BV-2 cells and the subsequent impact of SCFAs supplementation on the expression of TLR4, IKKα, and p65, as detected by immunoblotting. (O) Presentation of protein expression level statistics, indicating the modulatory effects of SCFAs on the TLR4/NF-κB pathway (n = 3). (P) Detection of TLR4, IKKα, and pp65 expression in BV-2 cells following blockade of the TLR4 pathway, further elucidating the mechanism of action (n = 3). (Q) Quantitative analysis of protein expression levels, reinforcing the role of TLR4/NF-κB signaling in the observed effects (n = 3). (R) Immunoblotting detection of TLR4, IKKα, and pp65 expression in brain tissues, providing in vivo evidence of the pathway’s involvement. (S) Quantitative analysis of protein expression density relative to the control group, underscoring the significance of TLR4/NF-κB modulation in the therapeutic effects (n = 3). Data are expressed as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, vs. Control group; #P < 0.05, ##P < 0.01, ###P < 0.001, vs. MCAO group (one- way analysis of variance followed by Tukey’s post hoc test). CCK-8: Cell counting kit-8; GAPDH: glyceraldehyde 3-phosphate dehydrogenase; IKKα: inhibitory kappa B kinase α; MCAO: middle cerebral artery occlusion; MnPB: manganese-iron Prussian blue; NF-κB: nuclear factor κB; RBITC: Rhodamine B isothiocyanate; SCFA: short-chain fatty acid; TLR4: Toll-like receptor 4.

Article Snippet: Cell culture and in vitro experimental design The human normal colonic epithelial cell line NCM460 (American Type Culture Collection, Manassas, VA, USA, CRL-1807, RRID:CVCL_2872) and BV-2 microglial cells (American Type Culture Collection, CRL-2467, RRID:CVCL_5744) were cultured in Dulbecco’s modified Eagle’s medium (Gibco, Carlsbad, CA, USA, Cat# 11965092) containing 10% fetal bovine serum (Thermo Fisher Scientific, Waltham, MA, USA, Cat# A5669701) and 1% of an antibiotic mixture containing penicillin/streptomycin (Gibco, Cat# 15140122).

Techniques: Concentration Assay, In Vitro, CCK-8 Assay, DCFH-DA Assay, Flow Cytometry, Expressing, Western Blot, In Vivo, Control, Cell Counting

Journal: BMC Cancer

Article Title: A novel machine learning-based immune prognostic signature for improving clinical outcomes and guiding therapy in colorectal cancer: an integrated bioinformatics and experimental study

doi: 10.1186/s12885-025-13437-0

Figure Lengend Snippet: Functional analysis of the representative gene APCDD1 in IRPS ( A ) Comparison of the APCDD1 expression level between CRC tissues and normal control tissues based on the GEPIA2 database; ( B ) The expression level of APCDD1 in CRC lines SW620, HT29 and SW480 was detected by qRT-PCR with the normal colon epithelial cell line NCM460 as a control; ( C ) The siRNA interference efficiency of APCDD1 in HT29 cell line was detected by qRT-PCR; ( D ) CCK-8 assays were used to detect the effect of APCDD1 knockdown on HT29 cell proliferation; ( E and F ) Transwell assays were used to detect the effect of APCDD1 knockdown on HT29 cell migration ( E ) and invasion ( F ); ( G ) Apoptosis assays were used to detect the effect of APCDD1 knockdown on apoptosis of HT29 cells. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: The human CRC cell lines HT29, SW480 and SW620, as well as the human normal colon mucosal epithelial cell line NCM460, were purchased from KeyGEN BioTECH Corp., Ltd (China).

Techniques: Functional Assay, Comparison, Expressing, Control, Quantitative RT-PCR, CCK-8 Assay, Knockdown, Migration