mab377 ab 2298772 mouse anti coup tf2 nr2f2 coup tf ii nr2f2  (R&D Systems)

Journal: Advanced Science

Article Title: Drug Repurposing for Treating NCOR‐Related Intellectual Disability

doi: 10.1002/advs.78030

Figure Lengend Snippet: NRG backward signaling contributes to NCOR1‐mediated regulation of the GABA‐A receptor. (A) Cell‐cell communication analysis showing upregulated NRG signaling in the NCOR1 KI/+ (nKI) vs. WT mice. (B) DEG analysis of upregulated gene expression of Nrg1/3 in the NCOR1 KI/+ vs. WT mice. (C) Nrg1 expression in the GABAergic and glutamatergic neurons. * p <0.05 by a two‐tailed t‐test. (D) Representative images of Neuro‐2a (N2a) cells transfected with empty vector (control), full‐length Nrg1 (NRG1‐FL), or NRG1‐ICD plasmids. (E) Western blot of transfected N2a cells with anti‐Flag antibody. β‐actin was the loading control. (F) RT‐qPCR analysis of transfected N2a cells with NRG1‐FL primers (left) targeting the non‐ICD region or ICD‐specific primers (right). (G) RT‐qPCR analysis of Gabra2 mRNA of transfected N2a cells. (H, I) Western blot and quantification of GABRA2 protein in N2a cells. (J, K) RT‐qPCR analysis of Nrg1 and Gabra2 mRNA after spironolactone (Spiro, 20 µM) treatment in N2a cells transfected with NRG1‐FL. (L, M) Western blot of GABRA2 protein in Spiro‐treated N2a cells transfected with NRG1‐FL. (N, O) RT‐qPCR analysis of Nrg1 and Gabra2 mRNA in Spiro‐treated N2a cells transfected with NRG1‐ICD. (P, Q) Western blot of GABRA2 protein in Spiro‐treated N2a cells transfected with NRG1‐ICD. n = 3 wells of cells for RT‐qPCR. n = 4 wells of cells for Western blot analysis. One‐way ANOVA and Tukey's post hoc test were used unless otherwise indicated. Data are mean ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: For Western blot, total cell lysates or brain tissue lysates were resolved by Tris‐glycine SDS‐PAGE, transferred to polyvinylidene difluoride (PVDF) membranes, and blotted with antibodies against Flag (F1804, MBL), β‐actin (81115‐1‐RR, Proteintech), Nrg1 (A0687, Abclonal), NCOR1 (A301‐146A, Bethyl), GAPDH (#2118L, CST), and Hsp90(4874S, CST).

Techniques: Analysis, Gene Expression, Expressing, Two Tailed Test, Transfection, Plasmid Preparation, Control, Western Blot, Quantitative RT-PCR

Journal: Advanced Science

Article Title: Drug Repurposing for Treating NCOR‐Related Intellectual Disability

doi: 10.1002/advs.78030

Figure Lengend Snippet: Pharmacologic targeting of NRG1 signaling improves cognition of NCOR1 KI/+ mice. NRG1‐ErbB antagonist spironolactone (Spiro, 50 mg/kg) or vehicle was administered systemically (i.p.) to 6‐month‐old female NCOR1KI/+ or WT mice daily for 3 days prior to the training day and 2 h before the behavioral training and testing sessions. (A, B) Discrimination index and total exploration time in the NOR test. (C, D) Discrimination index and total exploration time in the OLR test. (E) Escape latency of the Morris water maze (MWM) test during the 5‐day training. * p < 0.05 between KI/vehicle and the other groups by two‐way repeated ANOVA and Tukey's post hoc test. (F) Time percentage spent in the target quadrant zone during the probe test. (G) Total distance traveled in the MWM test. (H, I) Novel arm preference and total travel distance in the Y‐maze test. Data were analyzed by a two‐tailed unpaired t‐test. (J, K) Discrimination index and total exploration time in the 3CH‐SM test. (L, M) Social preference index and total exploration time in the 3CH‐SI test. Data were analyzed by a two‐tailed unpaired t‐test. (N) The ratio of time in open arms to time in closed arms during the EPM test. (O) Trace of exploration activity in the EPM test. (P) Time spent in the central area during the OFA test. (Q) Total travel distance during the OFA test. (R, S) Number of buried marbles during the test period. (T) RT‐qPCR analysis of the effects of Spiro on Gabra2 expression in the hypothalamus from WT and NCOR1 KI/+ (KI/+) mice. The sample size is indicated by individual data points. Each data point is one mouse (n = 7–19 per group). Data are mean ± SEM. Two‐way ANOVA, or two‐way repeated ANOVA followed by Tukey's post hoc test, was used unless otherwise indicated. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: For Western blot, total cell lysates or brain tissue lysates were resolved by Tris‐glycine SDS‐PAGE, transferred to polyvinylidene difluoride (PVDF) membranes, and blotted with antibodies against Flag (F1804, MBL), β‐actin (81115‐1‐RR, Proteintech), Nrg1 (A0687, Abclonal), NCOR1 (A301‐146A, Bethyl), GAPDH (#2118L, CST), and Hsp90(4874S, CST).

Techniques: Two Tailed Test, Activity Assay, Quantitative RT-PCR, Analysis, Expressing

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: NRG1 expression is elevated in RAW264.7 macrophages co-cultured with 4T1 cells. (A) The expression of NRG1 was examined using next-generation RNA sequencing analysis. (B) The mRNA expression of NRG1 in RAW264.7 macrophages co-cultured with 4T1 cells was detected using reverse transcription-quantitative PCR. (C) The protein expression of NRG1 in RAW264.7 macrophages co-cultured with 4T1 cells was detected using immunoblotting analysis. *** P<0.001. NRG1, neuregulin 1.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Cell Culture, RNA Sequencing, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: NRG1 expression is associated with M2 polarization and poor prognosis. (A) The expression of NRG1 in BC samples using TCGA database. (B) Spearman correlation analysis between NRG1 and CD163 expression in breast cancer samples. (C) The association of NRG1 expression with the overall survival in BC patients. NRG1, neuregulin 1; BC, breast cancer; TCGA, The Cancer Genome Atlas; GEPIA, Gene Expression Profiling Interactive Analysis.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Gene Expression

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Hypoxic MDA-MB-231 cells induces the M2 polarization of THP-1 macrophages via NRG1. (A) The mRNA expression of NRG1 was detected using RT-qPCR. (B) The transfection efficacy of Ov-NRG1 was detected using RT-qPCR and immunoblotting analysis. (C) The transfection efficacy of si-NRG1 was detected using RT-qPCR and immunoblotting analysis. (D) The level of IL-10 was detected using ELISA-related IL-10 assay kits. (E and F) The level of CD163 was detected using flow cytometry. (G) Following the treatment of anti-NRG1 blocking antibody, the level of IL-10 was detected using ELISA-related IL-10 assay kits. (H-I) Following the treatment of anti-NRG1 blocking antibody, the level of CD163 was detected using flow cytometry. * P<0.05, ** P<0.01 and *** P<0.001. NRG1, neuregulin 1; RT-qPCR, reverse transcription-quantitative PCR; Ov, overexpression; si, small interfering.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Blocking Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Over Expression

Journal: International Journal of Oncology

Article Title: Hypoxia-induced exosomal CAMTA1 promotes radio-resistance in MDA-MB-231 cells by regulating NRG1 to mediate M2 macrophage polarization

doi: 10.3892/ijo.2026.5875

Figure Lengend Snippet: Exosomal CAMTA1 promotes tumor growth in vivo . (A) The transfection efficacy of Ov-CAMTA1 was detected using RT-qPCR and immunoblotting analysis. (B) The appearance of tumor. The tumor (C) volume and (D) weight. (E) The mRNA expression of CAMTA1 was detected using RT-qPCR. (F) The level of IL-10 was detected using ELISA-related IL-10 assay kits. (G) The level of CD163 was detected using immunohistochemistry analysis. (H) H&E staining. (I) The expression of Caspase 3 was detected using immunohistochemistry analysis. (J) The Spearman correlation analysis of CAMTA1 and NRG1. (K) The expression of NRG1 was detected using immunohistochemistry analysis. * P<0.05, ** P<0.01 and *** P<0.001. CAMTA1, Calmodulin-binding Transcription Activator 1; Ov, overexpression; RT-qPCR, reverse transcription-quantitative PCR; H&E, hematoxylin and eosin; NRG1, neuregulin 1.

Article Snippet: Blocked by 5% BSA (MilliporeSigma) for 1 h at room temperature, the membranes were incubated with primary antibodies against NRG1 (cat. no. 10527-1-AP; 1:1,000; Proteintech Group, Inc.), hypoxia-inducible factor-1α (HIF-1α; cat. no. 20960-1-AP; 1:2,000; Proteintech Group, Inc.), CD63 (cat. no. 25682-1-AP; 1:2,000; Proteintech Group, Inc.), CD81 (cat. no. 27855-1-AP; 1:1,000; Proteintech Group, Inc.), CD9 (cat. no. 20597-1-AP; 1:2,000; Proteintech Group, Inc.), ALIX (cat. no. 12422-1-AP; 1:5,000; Proteintech Group, Inc.) and GAPDH (cat. no. 10494-1-AP; 1:5,000; Proteintech Group, Inc.) overnight at 4°C.

Techniques: In Vivo, Transfection, Quantitative RT-PCR, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Binding Assay, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction