Journal: Chinese Medicine

Article Title: Critical role of high-dose Astragalus in Buyang Huanwu Decoction for enhancing neurovascular coupling in a Qi Deficiency and Blood Stasis animal model

doi: 10.1186/s13020-026-01443-8

Figure Lengend Snippet: BHD HA promotes astrocyte development and enhances vascular coverage in CCI QDBS mice. A Representative immunofluorescence images of astrocytes (GFAP, green) and blood vessels (TRITC-Dextran, red) in the cortex of mice, bar = 100 μm; B Quantification of the total number of astrocytes; C Percentage of cerebrovascular-associated astrocytes; D Percentage of blood vessels covered with astrocytes; E Heatmap illustrating the expression levels of genes related to glial cell development in the Model, BHD HA, and BHD LA groups; F Representative immunofluorescence images of astrocytic NFIA in the cortex, bar = 100 μm; G ) Quantification of the mean fluorescence intensity of NFIA; H Quantification of the number of NFIA + /GFAP + cells, Data are presented as mean ± SEM. n = 3. ## p < 0.01, ### p < 0.001 vs. Sham; ** p < 0.01, *** p < 0.001 vs. Model; & p < 0.05, && p < 0.01, &&& p < 0.001 vs. BHD HA

Article Snippet: Ginaton (Dr. Willmar Schwabe GmbH & Co. KG); Isoflurane anaesthetic (RWD Life Science Co.,Ltd, R510-22-10); GFAP Antibody (Dako, M0761); NFIA Antibody (ABclonal, A3258); Donkey Anti-rabbit Alexa Fluor 594 (Invitrogen, A21207); Donkey Anti-Mouse Alexa Fluor 488 (Invitrogen, A 21202); pAAV-hSyn-jGCaMP7s-WPRE and pAAV-CMV-Luc2-WPRE-U6-shRNA( Nfia ) adeno-associated virus (OBiO Technology); Agarose Gel (Invitrogen, 16520050); Dental Casting Material (Shanghai New Century Dental Materials Co., Ltd.); TRITC-Dextran 155 kDa (Sigma, T1287); KW-BD Sleep Deprivation Device (Nanjing Calvin Biotechnology Co., Ltd.); Animal Anesthesia Machine (MATRX); High-Resolution Camera (Guangzhou Bohan Electronics Co., Ltd.); Ultrasonic Cleaner (Shanghai Zhixin Instrument Co., Ltd.); Grip Testing Instrument (Jinan Yiyan Technology Development Co., Ltd.); PharmaScan 70/16 US MRI spectrometer (Bruker, Germany); Stereotaxic Injection Apparatus (RWD Life Science Co.,Ltd); Cytation C10 (Biotek); Freeze Dryer (Shanghai Licheng Technology Co., Ltd.); Two-Photon Microscope (Olympus, Japan).

Techniques: Immunofluorescence, Expressing, Fluorescence

Journal: Chinese Medicine

Article Title: Critical role of high-dose Astragalus in Buyang Huanwu Decoction for enhancing neurovascular coupling in a Qi Deficiency and Blood Stasis animal model

doi: 10.1186/s13020-026-01443-8

Figure Lengend Snippet: NFIA knockdown reverses the beneficial effects of BHD HA on NVC and spatial memory. A Schematic diagram of AAV mediated Nfia knockdown. B confocal images of cortical sections stained for NFIA, the astrocyte marker GFAP, and blood vessels (TRITC-Dextran 155), bar = 50 μm. C Quantification of NFIA mean fluorescence intensity. D Quantification of the number of NFIA + /GFAP + cells. E Quantification of the total number of astrocytes; F Percentage of CAAs. G Percentage of blood vessels covered with astrocytes. n = 3. H Schematic diagram of NVC test using two-photon microscopy. I Representative images illustrating microvascular diameter before and after whisker stimulation. J Dilation analysis following whisker stimulation. K Representative images of XT line-scan. L Percentage change in microvascular blood flow following whisker stimulation. n = 5. M Representative movement trajectories of Barnes maze. N Proportion of effective exploratory activity in the target quadrant on day 4 (exploration time/total time in target quadrant). n = 6. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Ginaton (Dr. Willmar Schwabe GmbH & Co. KG); Isoflurane anaesthetic (RWD Life Science Co.,Ltd, R510-22-10); GFAP Antibody (Dako, M0761); NFIA Antibody (ABclonal, A3258); Donkey Anti-rabbit Alexa Fluor 594 (Invitrogen, A21207); Donkey Anti-Mouse Alexa Fluor 488 (Invitrogen, A 21202); pAAV-hSyn-jGCaMP7s-WPRE and pAAV-CMV-Luc2-WPRE-U6-shRNA( Nfia ) adeno-associated virus (OBiO Technology); Agarose Gel (Invitrogen, 16520050); Dental Casting Material (Shanghai New Century Dental Materials Co., Ltd.); TRITC-Dextran 155 kDa (Sigma, T1287); KW-BD Sleep Deprivation Device (Nanjing Calvin Biotechnology Co., Ltd.); Animal Anesthesia Machine (MATRX); High-Resolution Camera (Guangzhou Bohan Electronics Co., Ltd.); Ultrasonic Cleaner (Shanghai Zhixin Instrument Co., Ltd.); Grip Testing Instrument (Jinan Yiyan Technology Development Co., Ltd.); PharmaScan 70/16 US MRI spectrometer (Bruker, Germany); Stereotaxic Injection Apparatus (RWD Life Science Co.,Ltd); Cytation C10 (Biotek); Freeze Dryer (Shanghai Licheng Technology Co., Ltd.); Two-Photon Microscope (Olympus, Japan).

Techniques: Knockdown, Staining, Marker, Fluorescence, Microscopy, Whisker Assay, Activity Assay

Journal: Frontiers in Oncology

Article Title: NFIA-dependent upregulation of SMC4 promotes metastasis and metabolic reprogramming in glioma

doi: 10.3389/fonc.2025.1624370

Figure Lengend Snippet: NFIA promotes SMC4 transcription through direct binding to its promoter region in glioma cells. (A, B) The WB assay was used to detect the effects of NFIA overexpression and NFIA knockdown in U-251MG and LN229 cells, as well as the expression level of SMC4. (C) Schematic diagram of wild-type (wt) and mutant (mut, with mutated NFIA binding site) sequences of the SMC4 promoter. (D) The wild-type and mutant sequences of the SMC4 promoter were cloned into the pGL4 vector, and co-transfected with the pRL-TK vector (Renilla luciferase internal reference) into LN229 cells. Compared with the empty vector control group, overexpression of NFIA significantly enhanced the luciferase activity in the wt, mut2, and mut3 groups ( P < 0.0001), while there was no significant change in the mut1 group ( P = 0.9111). (E) The ChIP assay validates direct binding of NFIA to the SMC4 promoter region. ChIP-qPCR analysis was performed in LN229 cells with NFIA overexpression or control vector. Enriched DNA fragments spanning the predicted NFIA binding sites (-354 to -349 bp) in the SMC4 promoter were quantified and normalized to input DNA. (F) The IHC staining results of human gliomas showed that the expression of NFIA and SMC4 exhibited the same trend.

Article Snippet: Western blotting (WB) and immunofuorescence were performed using antibodies against NFIA (CST, #69375, Ameica), LDHA (proteintech, 19987-1-AP, China), SMC4 (proteintech, 24758-1-AP, China), GAPDH (affinity, AF7021, China), Smad2/3 (affinity, AF6367, China), p-Smad2/3 (affinity, AF3367, China), E-Cadherin (affinity, BF0219, China), N-Cadherin (affinity, AF5239, China), SNAI1 (affinity, AF6032, China), ZEB1 (affinity, DF7414, China) as primary antibodies.

Techniques: Binding Assay, Over Expression, Knockdown, Expressing, Mutagenesis, Clone Assay, Plasmid Preparation, Transfection, Luciferase, Control, Activity Assay, ChIP-qPCR, Immunohistochemistry

Journal: Science Advances

Article Title: Spinal motor neuron development and metabolism are transcriptionally regulated by nuclear factor IA

doi: 10.1126/sciadv.adu3346

Figure Lengend Snippet: ( A ) Volcano plot depicting differential gene expression between control (CT) and NFIA conditional knockout (NFIAcKO) ventral spinal cords ( n = 3). ( B ) Gene ontology (GO) analysis of DEGs in CT versus NFIAcKO falling under the categories of biological process, molecular function, and cellular component. GO analysis was performed separately on up- and down-regulated genes. ( C to F ) Heatmaps and hierarchical clustering of top 20 genes from GO categories mitochondrion organization (C), cytoplasmic translation (D), neuron projection (E), and neuron development (F), showing differences between CT and NFIAcKO gene expression. ( G ) NFIA ChIP-seq signals at NFIA-bound genomic regions in CT whole spinal cords. ( H ) Genomic locations of NFIA binding determined by ChIP-seq. ( I ) ChIP-qPCR analysis for Cdh2 , Ephb1 , and Epha4 using antibodies against NFIA or mouse control IgG in CT spinal cords ( n = 3). All data are presented as mean ± SEM. ** P ≤ 0.01 by Student’s t test.

Article Snippet: Genomic DNA regions of interest were isolated using 30 μl of rabbit anti-NFIA antibody (Atlas Antibodies).

Techniques: Gene Expression, Control, Knock-Out, ChIP-sequencing, Binding Assay, ChIP-qPCR