Journal: Signal Transduction and Targeted Therapy

Article Title: A biosafe mouse model for SARS-CoV-2 infection that more realistically simulates COVID-19 symptoms

doi: 10.1038/s41392-026-02640-5

Figure Lengend Snippet: SA-N-hACE2 mice are susceptible to SARS-CoV-2 ΔN/GFP-HiBiT infection. a Experimental design for intranasal infection. After TAM treatment, SA-N-hACE2 mice were infected either with 5 × 10 4 or 1 × 10 6 TCID 50 of SARS-CoV-2 ΔN/GFP-HiBiT. Tissue samples were collected at the indicated dpi. b Changes in the weights of the mice are shown ( n = 4 per group). c qRT‒PCR was used to quantify viral loads in tissues at 7 dpi ( n = 4 per group). The viral loads ( d ) and luminescence ( e ) were measured in the lungs collected at 2, 4, and 7 dpi ( n = 4 per group). f Immunofluorescence staining of lung sections at 2, 4, and 7 dpi was conducted using DAPI (blue) and an anti-GFP antibody (green) specific to GFP in SARS-CoV-2 ΔN/GFP-HiBiT. g – j Pathological changes observed using H&E staining in lung ( g ) and brain ( i ) tissues from SA-N-hACE2 mice challenged with 1 × 10 6 TCID 50 at 0, 2, 4, and 7 dpi. Pathology scores for the lungs ( h ) and brain ( j ) were calculated ( n = 4 per group). Data are presented as means ± SD. Statistical significance was determined using one-way ANOVA with Dunnett’s multiple comparisons test ( h ). ns not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ND not detected

Article Snippet: The samples were first immunostained with an anti-GFP antibody (Sino Biological, 13105-R208, 1:500) followed by an Alexa Fluor 488-conjugated secondary antibody (Thermo Fisher Scientific, A-11008, 1:1000).

Techniques: Infection, Immunofluorescence, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: A biosafe mouse model for SARS-CoV-2 infection that more realistically simulates COVID-19 symptoms

doi: 10.1038/s41392-026-02640-5

Figure Lengend Snippet: Sftpc-N-hACE2 mice are susceptible to SARS-CoV-2 ΔN/GFP-HiBiT infection. a Illustration of the protocol for intranasal infection with tissue samples collected at the indicated dpi. Sftpc-N-hACE2 mice were infected with 5 × 10 4 or 1 × 10 6 TCID 50 of SARS-CoV-2 ΔN/GFP-HiBiT. b Changes in the body weights of the mice are shown ( n = 4 per group). c Viral loads in the tissues obtained at 7 dpi were quantified via qRT‒PCR ( n = 4 per group). The viral loads ( d ) and luminescence ( e ) were measured in the lungs collected at 2, 4, and 7 dpi ( n = 4 per group). f Immunofluorescence staining of lung sections at 2, 4, and 7 dpi was conducted using DAPI (blue) and an anti-GFP antibody (green) specific to GFP in SARS-CoV-2 ΔN/GFP-HiBiT. g – j Pathological changes in the lungs ( g ) and brains ( i ) of Sftpc-N-hACE2 mice challenged with 1 × 10 6 TCID 50 at 0, 2, 4, and 7 dpi were assessed using H&E staining. Pathology scores for the lungs ( h ) and brains ( j ) are shown ( n = 4 per group). Data are presented as means ± SD. Statistical significance was determined using one-way ANOVA with Dunnett’s multiple comparisons test ( h ). ns not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ND not detected

Article Snippet: The samples were first immunostained with an anti-GFP antibody (Sino Biological, 13105-R208, 1:500) followed by an Alexa Fluor 488-conjugated secondary antibody (Thermo Fisher Scientific, A-11008, 1:1000).

Techniques: Infection, Immunofluorescence, Staining

Journal: Signal Transduction and Targeted Therapy

Article Title: A biosafe mouse model for SARS-CoV-2 infection that more realistically simulates COVID-19 symptoms

doi: 10.1038/s41392-026-02640-5

Figure Lengend Snippet: SARS-CoV-2 ΔN/GFP-HiBiT infection in Sftpc-N-hACE2 mice can lead to lethal disease. a The schematic outlines the procedure for infecting the mice with 5 × 10 6 TCID 50 of SARS-CoV-2 ΔN/GFP-HiBiT. At 0, 7, 10, 14, and 21 dpi, four mice were euthanized at each time point for sample collection. Mice that experienced a loss of more than 20% of their initial body weight were euthanized as a humane endpoint. Mice were monitored for body weight changes ( b ) and survival ( c ) ( n = 16). E gene copies in lung ( d ) and brain ( e ) tissues were quantified via qRT‒PCR. f – i Pathological changes were assessed using H&E staining in the lungs ( f ) and brains ( g ). Pathology scores for the lungs ( h ) and brains ( i ) were recorded ( n = 4 per group). j Immunofluorescence staining of brain sections was performed using DAPI (blue) and an anti-GFP antibody (green) specific to GFP in SARS-CoV-2 ΔN/GFP-HiBiT. k Immunofluorescence analysis was performed on brain sections from mock-infected mice, K18-hACE2 KI mice challenged with live SARS-CoV-2, and both SA-N-hACE2 and Sftpc-N-hACE2 mice exposed to SARS-CoV-2 ΔN/GFP-HiBiT. The sections were stained with anti-IBA1 (red) and anti-CD68 (green) antibodies. l Light-sheet imaging of cleared lung tissues from Sftpc-N-hACE2 mice at 7 dpi confirmed the distribution of SARS-CoV-2 N in lung epithelial cells and SARS-CoV-2 ΔN/GFP-HiBiT infection. Data are presented as means ± SD. Statistical significance was determined using one-way ANOVA with Dunnett’s multiple comparisons test ( h ). ns not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. ND not detected

Article Snippet: The samples were first immunostained with an anti-GFP antibody (Sino Biological, 13105-R208, 1:500) followed by an Alexa Fluor 488-conjugated secondary antibody (Thermo Fisher Scientific, A-11008, 1:1000).

Techniques: Infection, Staining, Immunofluorescence, Imaging

Journal: Biochemical and biophysical research communications

Article Title: Metal-metal interaction mediates the iron induction of Drosophila MtnB.

doi: 10.1016/j.bbrc.2017.04.109

Figure Lengend Snippet: Fig. 2. MtnB expression is regulated by iron in Drosophila. A. RT-PCR of MtnB, MtnC and MtnD under different iron conditions. RNAs were made from the whole body of third instar larvae. B. MtnB expression at the protein level. MtnB-EGFP flies were adopted to examine iron influence on MtnB expression. Anti-GFP was used as the primary antibody in the western blot. Total protein came from the whole body of the adult flies. C. Fluorescence of MtnB-EGFP in the anterior midgut. D. MtnB-EGFP expression in the posterior midgut. Fruit flies were raised on cornmeal-agar food containing either iron, zinc or copper. Midguts were from third instar larvae.

Article Snippet: Anti-GFP mouse monoclonal antibody was from Zhongshan Goldenbridge Biotechnology (Beijing, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Fluorescence

Journal: Frontiers in Cellular Neuroscience

Article Title: Mob2 Insufficiency Disrupts Neuronal Migration in the Developing Cortex

doi: 10.3389/fncel.2018.00057

Figure Lengend Snippet: Biallelic variants in MOB2 affect mRNA and protein stability in vitro . (A) Brain MRI scan of patient 1203, showing the presence of bilateral periventricular nodular heterotopia (PH; red arrows). (B) Sequence traces illustrating the MOB2 sequence variants c.679G > A, present in mother and patient, and c.207delC, present in father and patient. (C) Multiple sequence alignment (ClustalX) of the human MOB2 protein against various species. Columns are colored by conservation and property. * Denotes the positions of the mutated residues—Phe69 and Glu227—substituted by each of the alleles discovered in individual 1203. (D) Mean expression of MOB2 in age-matched controls and individual 1203 ( n = 5) fibroblasts when treated with vehicle/DMSO (black circles) and cycloheximide (CHX; red squares), as determined by real-time PCR. Expression was normalized against GAPDH and DIMT1 housekeeping genes in the same sample using the relative standard curve method. Values were then subsequently normalized to control 1 DMSO treatment using the delta-delta Ct method. (E) HEK293 cells were transiently transfected with a bi-cistronic vector expressing both the green fluorescent protein (GFP) and wild-type (w.t) or mutant (Glu227Lys) MOB2 . At 24 h after transfection, cells were treated with DMSO, or the proteasome inhibitor, MG132, for 24 h, after which total cell lysates were prepared and subjected to western blot analysis using anti-MOB2, anti-GFP and anti-GAPDH as a loading control. (F) Quantifications representing three biological replicates of (E) summarizing total MOB2 protein amount with or without MG132 treatment. Lysosomal degradation pathways may account for the remaining degraded MOB2. ** P < 0.01; *** P < 0.001, Mann-Whitney U test.

Article Snippet: Media containing DMEM, 10% FCS, 1% L-glutamine and either solvent alone (0.01% DMSO) or the proteasome inhibitor, 1 μM MG132 (Sigma-Aldrich, Cat. # C2211), in DMSO for 24 h. After which point total cell lysates were prepared and subjected to Western blot analysis using MOB2 (mouse, LSBio, Cat. #LS-C184588, 1:5000), GFP (mouse, OriGene, Cat. # TZ150041, 1:5000) and GAPDH (rabbit, Sigma-Aldrich, Cat. # G9545, 1:3000).

Techniques: In Vitro, Sequencing, Expressing, Real-time Polymerase Chain Reaction, Control, Transfection, Plasmid Preparation, Mutagenesis, Western Blot, MANN-WHITNEY

Journal: Frontiers in Cellular Neuroscience

Article Title: Mob2 Insufficiency Disrupts Neuronal Migration in the Developing Cortex

doi: 10.3389/fncel.2018.00057

Figure Lengend Snippet: Mob2 knockdown alters neuronal cell distribution and nucleus-cilia coupling. Coronal micrograph sections of E16 mouse cerebral cortices electroporated at E13 with EGFP/empty vector control (A) or miRNAs targeting Mob2 (B) . (C) Quantification of the distribution of EGFP-expressing (EGFP+) cells transfected with EGFP/empty vector alone or miRNAs targeting Mob2 3 days after electroporation (mean ± SEM). The cortex was subdivided into five equal width bins approximately corresponding to VZ (bin 1), SVZ (bin 2), IZ (bin 3) and CP (bins 4 and 5). CP, cortical plate; IZ, intermediate zone; SVZ, subventricular zone; VZ, ventricular zone; dpe, days post electroporation. At least three embryos were analyzed for each condition. n, total number of GFP+ cells counted per condition. (D,E) Representative images of migrating neurons in E16 cortices (3 dpe) electroporated with EGFP/control or miRNA targeting Mob2 (green) and Arl13b-tagRFP (red) located within bin 5 (CP2). Arrow heads indicate the cell bodies and arrows indicate the cilia labeled by Arl13b-tagRFP. Asterisks show the position of three cilia within a neuron. (F) Quantification of the distance between the cilia and nucleus (micrometers) in neurons. Each dot represents a neuron and red lines indicate mean ± SEM. (G) Quantification of the number of cilia in each neuron. (H,I) Micrographs sections of day 51 human cerebral organoids electroporated at day 44 with EGFP/empty vector control or miRNA targeting MOB2 (green) and Arl13b-tagRFP (red). White arrows indicate the cilia labeled by Arl13b-tagRFP. (J) Quantification of the percentage of cells with more than one cilium of organoids transfected with EGFP/empty vector control (GFP) or miRNAs targeting Mob2 . Data taken from at least three ventricular structures. (C,F,G) Mann-Whitney U test, (J) Exact binomial test; * p < 0.05, *** p < 0.001. Scale bar represents (A,B) 100 μm, (D,E) 10 μm, (H,I) 25 μm and (H′,H″,I′,I″) 5 μm.

Article Snippet: Media containing DMEM, 10% FCS, 1% L-glutamine and either solvent alone (0.01% DMSO) or the proteasome inhibitor, 1 μM MG132 (Sigma-Aldrich, Cat. # C2211), in DMSO for 24 h. After which point total cell lysates were prepared and subjected to Western blot analysis using MOB2 (mouse, LSBio, Cat. #LS-C184588, 1:5000), GFP (mouse, OriGene, Cat. # TZ150041, 1:5000) and GAPDH (rabbit, Sigma-Aldrich, Cat. # G9545, 1:3000).

Techniques: Knockdown, Plasmid Preparation, Control, Expressing, Transfection, Electroporation, Labeling, MANN-WHITNEY