neuro 2a n2a mouse neuroblastoma cell lines  (ATCC)

Journal: Alzheimer's & Dementia

Article Title: NRN1 as a therapeutic target for Alzheimer's disease

doi: 10.1002/alz.71149

Figure Lengend Snippet: Abcam ab64186 recognizes NRN1 protein in mouse neuroblastoma cells. (A) Representative Western blot of human embryonic kidney (HEK) 293T, Neuro‐2a (N2a) mouse neuroblastoma, and SH‐SY5Y human neuroblastoma cell lysates (50 µg protein), probed with NRN1 polyclonal antibody Abcam ab64186 and GAPDH monoclonal antibody. (B) N2a cells were transfected with NRN1 or Scramble (non‐targeting) siRNA smart pools and harvested after 96 h. Western blot analyses with NRN1 polyclonal antibody Abcam ab64186 revealed reduced intensity of the ∼34 kDa band in NRN1‐depleted cells, compared to Scramble control. 10 µg of protein was loaded per lane, and GAPDH was probed as a loading control. GAPDH, glyceraldehyde‐3‐phosphate dehydrogenase; NRN1, Neuritin‐1; siRNA, small interfering RNA;

Article Snippet: Neuro‐2a (N2a) mouse neuroblastoma cell lines (Catalog No.: CCL‐131, ATCC) were maintained in MEM (Catalog No.: 11095‐080, Thermo Fisher Scientific) with 10% FBS and 1% penicillin‐streptomycin.

Techniques: Western Blot, Transfection, Control, Small Interfering RNA

Journal: Cell Death & Disease

Article Title: Targeting ENO1 reprograms macrophage polarization to trigger antitumor immunity and improves the therapeutic effect of radiotherapy

doi: 10.1038/s41419-026-08416-7

Figure Lengend Snippet: A A total of 2.5 × 10 5 CT26 cells were subcutaneously injected into BALB/c mice for 11 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 20 mg/kg) five times at three-day intervals ( n = 5). The tumor volume was recorded every three days. * p < 0.05. Two-Way ANOVA test ( n = 5). B The resected tumors from the CT26 tumor-bearing mice were analyzed by immunohistochemical analysis. ** p < 0.01. Unpaired t test ( n = 5). C A total of 2.5 × 10 5 CT26 cells were intravenously injected into BALB/c mice for 11 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 40 mg/kg) five times at three-day intervals ( n = 5). The tumor volume was recorded every three days. The resected lung tissues were analyzed by HE staining. *** p < 0.001. Unpaired t test ( n = 5). D 4T1 cells (5 × 105) were intravenously injected into BALB/c mice, which were then intraperitoneally administered anti-ENO1 antibodies (HuL001, 40 mg/kg) on the indicated days (n = 5). The tumor volume was recorded every three days. * p < 0.05. Two-Way ANOVA test ( n = 5). E The resected tumors from 4T1-bearing mice were analyzed by HE staining. ** p < 0.01. Unpaired t test ( n = 5). F The resected lung tissues from 4T1-bearing mice were analyzed by HE staining, immunofluorescence staining and immunohistochemical analysis. ** p < 0.01 and *** p < 0.001. Unpaired t test ( n = 5). G The resected liver tissues from 4T1-bearing mice were analyzed by HE staining, immunofluorescence staining and immunohistochemical analysis. *** p < 0.001. Unpaired t test ( n = 5). H The density of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors from CT26- and 4T1-bearing mice was evaluated by immunofluorescence staining ( n = 3). I The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors from CT26-bearing mice. * p < 0.05 and *** p < 0.001. Unpaired t test ( n = 5). J The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors from 4T1-bearing mice. * p < 0.05 and *** p < 0.001. Unpaired t test ( n = 5). K The DEGs between HuL001- and Veh-treated resected CT26 tumors are shown in a volcano plot ( n = 2). L Compared with those in Veh.-treated resected tumors, the number of glycolysis-related gene sets significantly decreased in HuL001-treated resected tumors ( n = 2).

Article Snippet: The mouse colon cancer cell line CT26, mouse breast cancer cell line 4T1, human CRC cell lines (SW480, SW620, HCT116, LoVo and HT29) and human breast cancer cell lines (MDA-MB-468, BT-20, HS578T and SUM-159) were obtained from the ATCC.

Techniques: Injection, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Cell Death & Disease

Article Title: Targeting ENO1 reprograms macrophage polarization to trigger antitumor immunity and improves the therapeutic effect of radiotherapy

doi: 10.1038/s41419-026-08416-7

Figure Lengend Snippet: A A total of 5 × 10 5 CT26 cells were subcutaneously injected into the left legs of BALB/c mice for 5 days and then intraperitoneally administered with anti-ENO1 antibodies (HuL001, 40 mg/kg) or clodronate liposomes (50 μL/mouse) on the indicated days ( n = 5). Local radiotherapy was given on Day 10. The tumor volume was recorded every three days. * p < 0.05. Two-Way ANOVA test ( n = 4). B The densities of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors were analyzed by immunofluorescence staining. The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors. * p < 0.05 and *** p < 0.001. One-Way ANOVA test ( n = 3). C 4T1 cells (5 ×10 4 ) were subcutaneously injected into the left legs of BALB/c mice for 4 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 20 mg/kg) six times on the indicated days ( n = 5). Local radiotherapy was given on Days 10 and 12. The tumor volume was recorded every three days. * p < 0.05 and *** p < 0.001. Two-Way ANOVA test ( n = 4). D The resected tumors were weighed on Day 40. * p < 0.05. One-Way ANOVA test ( n = 4). E The densities of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors were analyzed by immunofluorescence staining. F The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors. * p < 0.05. One-Way ANOVA test ( n = 3). G The frequencies of M1 (CD11c + CD11b + F4/80 + CD45 + 7AAD - CD3 - CD19 - ) and M2 (CD1206 + CD11b + F4/80 + CD45 + 7AAD - CD3 - CD19 - ) tumor-infiltrating macrophages were analyzed by flow cytometry. * p < 0.05. One-Way ANOVA test ( n = 3-4). H The quantification of the M1/M2 ratio is shown. * p < 0.05. One-Way ANOVA test ( n = 3). I A representative image of flow cytometric analysis of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells. J The frequency of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells was analyzed by flow cytometry. * p < 0.05. One-Way ANOVA test ( n = 3-4). K The density of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells is shown. * p < 0.05. One-Way ANOVA test ( n = 3-4). L The density of IFNγ + (IFNγ hi CD8 + CD3 + CD45 + 7AAD - ) T cells is shown ( n = 3-4). * p < 0.05 and ** p < 0.01. One-Way ANOVA test ( n = 3-4). M The proposed mechanism of TGFβ1/TGFβR/Smad3/PRMT5-mediated ENO1 translocation for lactate release via MCT4.

Article Snippet: The mouse colon cancer cell line CT26, mouse breast cancer cell line 4T1, human CRC cell lines (SW480, SW620, HCT116, LoVo and HT29) and human breast cancer cell lines (MDA-MB-468, BT-20, HS578T and SUM-159) were obtained from the ATCC.

Techniques: Injection, Liposomes, Immunofluorescence, Staining, Flow Cytometry, Translocation Assay

Journal: Redox Biology

Article Title: Combination of YAP inhibition and photodynamic therapy induces dual DNA damage and activates STING pathway to enhance immunotherapy in uveal melanoma

doi: 10.1016/j.redox.2025.103965

Figure Lengend Snippet: Activation of the cGAS-STING pathway by HANP/VP via YAP-inhibition and PDT in vitro. (A) The phosphorylation levels of representative proteins of the cGAS-STING pathway were analyzed using Western blot ( left ) and quantified ( right ). After HANP/VP treatment without laser irradiation, the pSTING/STING, pTBK1/TBK1, and pIRF3/IRF3 ratio were significantly increased compared to the untreated cells; the phosphorylation levels were further increased after laser irradiation, n = 3/group. (B) qPCR was used to detect IFN-β1 mRNA levels. Compared with the control group, HANP/VP without Laser treatment significantly increased IFN-β1 expression in B16F10 cells, while HANP/VP combined with laser irradiation further elevated IFN-β1 mRNA levels. When cells were pretreated with ethidium bromide (EB, 120 ng/mL), IFN-β1 mRNA levels decreased in the HANP/VP with Laser irradiation group. Following H151 treatment, IFN-β1 mRNA levels decreased to control group levels regardless of laser irradiation status. n = 3/group. Statistical significance was determined by one-way ANOVA (∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: The mouse melanoma cell line B16F10 was purchased from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Activation Assay, Inhibition, In Vitro, Phospho-proteomics, Western Blot, Irradiation, Control, Expressing

Journal: Glycobiology

Article Title: Editor's Choice Functional inactivation of oligosaccharyltransferase a isoform suppresses tumor metastasis

doi: 10.1093/glycob/cwag003

Figure Lengend Snippet: Syngeneic model. (A) Schematic representation of the experimental model. When cancer cell-derived EVs that contain Cre25nt mRNA were incorporated into recipient cells, the mRNA may be translated to Cre recombinase, leading to expression of tdTomato protein. (B and C) RT-PCR and western blot analysis of Cre25nt mRNA (B) and Cre protein (C) in wild-type (WT) B16-F10 cells, Cre25nt cells, and small EVs secreted from these cells. (D) Expression of tdTomato (red) in tumors formed by WT cells and Cre25nt cells. tdTomato protein was also detected by anti-red fluorescent protein (RFP) antibody (green). Sections were counterstained with Hoechst 33342 (cyan). Scale bars, 100 μm.

Article Snippet: The mouse melanoma cell line B16-F10 was purchased from American Type Culture Collection (ATCC).

Techniques: Derivative Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: Glycobiology

Article Title: Editor's Choice Functional inactivation of oligosaccharyltransferase a isoform suppresses tumor metastasis

doi: 10.1093/glycob/cwag003

Figure Lengend Snippet: Gene-edited knockout of the Stt3a and Stt3b genes in mouse melanoma cells. (A and B) Western blot analysis of wild-type (WT) B16-F10, Stt3a -RE, Stt3a -KO, Stt3b -RE, and Stt3b -KO cells. Samples were treated with or without PNGase F (B). (C) Quantitative real-time PCR analysis of Cre25nt mRNA in small EVs secreted from Stt3a -RE, Stt3a -KO, Stt3b -RE, and Stt3b -KO cells. Average expression of Stt3a -KO, Stt3b -KO were set to 1.0. Data represent the mean ± standard deviation; n = 3.

Article Snippet: The mouse melanoma cell line B16-F10 was purchased from American Type Culture Collection (ATCC).

Techniques: Knock-Out, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation