Journal: PLOS Pathogens

Article Title: CNP blocks mitochondrial depolarization and inhibits SARS-CoV-2 replication in vitro and in vivo

doi: 10.1371/journal.ppat.1011870

Figure Lengend Snippet: ( a ) Groups of mice were transduced intranasally with rAd5 vectors (1 x 10 8 viral particles/ mouse) dually expressing hACE2 and either HA-tagged CNP or HA-tagged eGFP 3 days prior to intranasal infection with SARS-CoV-2/WA1 (1 x 10 3 or 1 x 10 5 PFU/mouse). Image created using BioRender. b - c Infectious viral loads from lung homogenates were determined by plaque assay at 2- or 4- days post SARS-CoV-2 challenge for the ( b ) 1 x 10 3 PFU/mouse or ( c ) 1 x 10 5 PFU/mouse challenge groups. d - e Virus titers were also determined by RT-qPCR for sgRNA for the ( d ) 1 x 10 3 PFU/mouse or ( e ) 1 x 10 5 PFU/mouse challenge groups. *** p ≤ 0.001, ****p ≤ 0.0001. Virus titer data is the shown for three experiments.

Article Snippet: Two mammalian gene expression recombinant Adenovirus serotype 5 (rAd5) viral vectors were prepared by and obtained from VectorBuilder Inc. to dually express hACE2 and either: (1) HA-tagged hCNP (Ad/CNP); or (2) HA-tagged enhanced green fluorescent protein (Ad/eGFP).

Techniques: Expressing, Infection, Plaque Assay, Virus, Quantitative RT-PCR

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Super enhancer-driven LINC01013 mediates hypoxia-induced mitochondrial dysfunction by HSPA9 to determine pulmonary arterial smooth muscle cell fate

doi: 10.1007/s00018-025-06071-3

Figure Lengend Snippet: Human LINC01013 delivery aggravates PH progression in mice. A - B Right ventricular systolic pressure (RVSP) and RV/left ventricular (LV) + Septum weight ratio in the SuHx-induced PH mouse models ( n = 6). C Pulmonary artery velocity time integral (PAVTI), pulmonary artery acceleration time (PAAT) and left ventricular ejection fraction (LVEF) of the SuHx-induced PH mice models infected with AAV5 carrying human LINC01013 ( n = 6). D Expression of PCNA in lung tissues ( n = 5). E Pulmonary arterial morphological analysis was performed by using hematoxylin and eosin (HE) and Masson staining. Scale bar, 100 μm. F-G Immunofluorescence of TNF-α, and activities of SOD and Gpx in lung tissues ( n = 6). Scale bars, 50 μm. H RNA pull-down assays detected the interaction between LINC01013 and HSPA9 protein in mouse PASMCs. All values are presented as the mean ± SEM. Statistical analysis was performed with one-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001. Nor, normoxia; SuHx, hypoxic + Su5416; NC, negative control; ▲SMC, smooth muscle cell targeting

Article Snippet: Clonal constructs of LINC01013 targeting the smooth muscle cell-specific promoter SM22α (smooth muscle 22α) were packaged into serotype 5 adeno-associated virus (AAV5) vectors and synthesized by GeneChem (Shanghai, China) (Fig. A).

Techniques: Infection, Expressing, Staining, Immunofluorescence, Negative Control

Journal: Cells

Article Title: Astrocytic CD44 Deficiency Reduces the Severity of Kainate-Induced Epilepsy

doi: 10.3390/cells12111483

Figure Lengend Snippet: GFAP-Cre recombinase viral approach to target CD44 in adult dentate gyrus astrocytes. ( a ) Scheme of bilateral intrahippocampal administration of AAV vectors (CTRL: AAV5. gfaABC1D ::GFP; GFP localizes in the cytoplasm of astrocytes, and AsKO: AAV5. gfaABC1D ::GFP-Cre; GFP-Cre localizes in cell nuclei of astrocytes) into the brains of CD44 fl/fl mice. ( b ) CD44 level in protein extracts from DG of CTRL and AsKO animals (Western blot). Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as the loading control protein. ( c ) Quantitative analysis of Western blot results ( n = 6 independent animals, Student’s t -test, *** p < 0.001, data presented as mean ± SEM) ( d ) CD44-immunoreactivity in astrocytic s100β profiles from the ML of the DG in CTRL and AsKO mice (see panel f for representative immunostaining) (Student’s t -test, *** p < 0.001, data presented as mean ± SEM). Representative immunofluorescence micrographs of GFP (green), astrocyte marker s100β (white), and CD44 (red) expression in the hippocampus of CTRL and AsKO mice for matching triple stain images shown at ( e ) a lower magnification or ( f ) the close-ups reflecting green squares on panel ( e ). Arrows point to astrocytes (s100β staining) that were or were not transfected (green arrow for GFP-expressing and white arrow for non-expressing, respectively). ML—molecular layer, GL—granular layer. Scale bar: ( e ) 200 μm and ( f ) 20 μm.

Article Snippet: To produce animals with the astrocytic knockout of CD44 (AsKO), one group was stereotaxically injected with 500 nL of AAV5 vector with Cre recombinase–AAV5. gfaABC1D ::GFP-Cre (construct: Bryan Roth, AAV Serotype 5, UNC Vector Core, Chapel Hill, NC, USA, AAV vector with ITR2 where the human gfaABC1D promoter [ ] drives the expression of eGFP attached to the N-terminus of Cre recombinase), whereas the control (CTRL) group was injected with 500 nL of AAV5. gfaABC1D ::GFP vector (Construct: Bryan Roth, Addgene plasmid #50473; AAV Serotype 5, UNC Vector Core, AAV vector with ITR2 where the human gfaABC1D promoter drives the expression of eGFP).

Techniques: Western Blot, Control, Immunostaining, Immunofluorescence, Marker, Expressing, Staining, Transfection

Journal: Cells

Article Title: Astrocytic CD44 Deficiency Reduces the Severity of Kainate-Induced Epilepsy

doi: 10.3390/cells12111483

Figure Lengend Snippet: CD44 deletion in astrocytes reduces reactive astrogliosis following kainate-induced epilepsy. Representative images of astrocytes (GFAP, white), CD44 expression (red), GFP (green), and nuclei (Hoechst, purple) in the hippocampal ML of the DG of CTRL and CD44 AsKO mice before ( a ) and after ( b ) 4 weeks post-KA administration. Scale bar: 20 μm. Quantitative analysis of GFAP-immunoreactive area (µm 2 ) in the ML of the DG in CTRL and CD44 AsKO mice 4 weeks after administration of ( c ) SA (Kolmogorov-Smirnov test, p = 0.41, data presented as median with IQR) or ( e ) KA ( n = 6 independent animals in each group, Student’s t -test with Welch’s correction, ** p = 0.004, data presented as mean ± SEM). Quantitative analysis of CD44-immunoreactivity in astrocytic GFAP profiles in the ML of the DG in CTRL and CD44 AsKO mice 4 weeks after administration of ( d ) SA (Student’s t -test with Welch’s correction, *** p < 0.001, data presented as mean ± SEM) or ( f ) KA (Student’s t -test with Welch’s correction, ** p = 0.001, data presented as mean ± SEM).

Article Snippet: To produce animals with the astrocytic knockout of CD44 (AsKO), one group was stereotaxically injected with 500 nL of AAV5 vector with Cre recombinase–AAV5. gfaABC1D ::GFP-Cre (construct: Bryan Roth, AAV Serotype 5, UNC Vector Core, Chapel Hill, NC, USA, AAV vector with ITR2 where the human gfaABC1D promoter [ ] drives the expression of eGFP attached to the N-terminus of Cre recombinase), whereas the control (CTRL) group was injected with 500 nL of AAV5. gfaABC1D ::GFP vector (Construct: Bryan Roth, Addgene plasmid #50473; AAV Serotype 5, UNC Vector Core, AAV vector with ITR2 where the human gfaABC1D promoter drives the expression of eGFP).

Techniques: Expressing

Journal: eLife

Article Title: Neural correlates and determinants of approach–avoidance conflict in the prelimbic prefrontal cortex

doi: 10.7554/eLife.74950

Figure Lengend Snippet:

Article Snippet: Recombinant DNA reagent , pAAV-mDlx-ChR2-mCherry-Fishell-3 , Addgene and University of North Carolina Viral Vector Core , Addgene plasmid #83,898 , Packaged by UNC vector core (Serotype 5).

Techniques: Recombinant, Plasmid Preparation, Inhibition, Control, Sequencing, RNAscope, Multiplex Assay, Software