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: Plant physiology and biochemistry : PPB

Article Title: Gibberellic acid signaling promotes resistance to saline-alkaline stress by increasing the uptake of ammonium in rice.

doi: 10.1016/j.plaphy.2024.108424

Figure Lengend Snippet: Fig. 4. The interaction of SLR1 with bZIP23 and the responses of bZIP23 plants to SAK stress. (a) A Y2H assay was performed to assess the interaction between BD- bZIP23 and AD-SLR1. Yeast cells transformed with the vectors indicated were grown on SD/-Trp -Leu or SD/-Trp -Leu -His + 10 mM and 15 mM 3-AT. EV: empty vector. (b) A BiFC assay was performed to analyze the interaction between bZIP23 and SLR1. The reconstruction of YFP from SLR1-cCFP + nYFP, cCFP + bZIP23- nYFP, or SLR1-cCFP + bZIP23-nYFP was imaged. H2B-RFP was used as the nuclear marker, and differential interference contrast (DIC) and merged images were captured. Bar = 20 μm. (c) A Co-IP assay was performed to assess the interaction between bZIP23-GFP and SLR1-Myc. Anti-GFP antiserum was used for immu noprecipitation, and anti-GFP or anti-Myc antibodies were used for a WB analysis. (d) (f) bzip23, bZIP23 OX and their corresponding WT plants were grown on 0.5X MS, 0.5X MS + 80 mM of NaHCO3 (SAK stress) for 10 days. (e) (g) The inhibition of primary root growth was calculated from the plants grown under SAK stress conditions. The experiments were repeated at least three times. Significant differences among more than two groups at P < 0.05 are marked with different letters. 3- AT, 3-aminotriazole.

Article Snippet: Western blotting was performed using the following primary antibodies: anti-GFP antibody (AE011, ABclonal Technology, Wuhan, China) and anti-Myc antibody (AE010, ABclonal Technology, Wuhan, China).

Techniques: Y2H Assay, Transformation Assay, Plasmid Preparation, Bimolecular Fluorescence Complementation Assay, Marker, Co-Immunoprecipitation Assay, Inhibition

Journal: Plant physiology and biochemistry : PPB

Article Title: Gibberellic acid signaling promotes resistance to saline-alkaline stress by increasing the uptake of ammonium in rice.

doi: 10.1016/j.plaphy.2024.108424

Figure Lengend Snippet: Fig. 5. Interaction between IDD10 and bZIP23 and their influence on gene activation. (a) A Y2H assay was performed to assess the interaction between BD-IDD10 and AD-bZIP23. Yeast cells transformed with the vectors indicated were grown on SD/-Trp -Leu or SD/-Trp -Leu -His. EV: empty vector. (b) A BiFC assay was performed to analyze the interaction between bZIP23 and SLR1. The reconstruction of YFP from cCFP + nYFP, IDD10-cCFP + nYFP, cCFP + bZIP23-nYFP, or IDD10- cCFP + bZIP23-nYFP was imaged. H2B-RFP was used as the nuclear marker, and differential interference contrast (DIC) and merged images were captured. Bar = 20 μm. (c) A Co-IP assay was performed to assess the interaction between bZIP23-GFP and IDD10-Myc. Anti-Myc antiserum was used for immunoprecipitation, and anti- GFP or anti-Myc antibodies were used for a WB analysis. (d) A transient expression assay was performed to analyze the effects of IDD10, bZIP23 and IDD10 + bZIP23 on the activation of the AMT1;2 promoter (pAMT1;2). The experiments were repeated at least three times. Significant differences among more than two groups at P < 0.05 are marked with different letters. (e) (f) Schematic diagram showing the position of bZIP23 binding in the AMT1;2 promoter of the probe used to detect ChIP. The gray oval represents bZIP23 binding, and the letters P (1, 2) indicate the positions. (g) NH4 + contents in the roots of WT, bZIP23 OX, and bzip23 plants were measured. (h) A transient expression assay was performed to analyze the effects of bZIP23 and bZIP23+SLR1 on the activation of AMT1;2 promoter (pAMT1;2). The experiments were repeated at least three times. Significant differences among more than two groups at P < 0.05 are marked with different letters.

Article Snippet: Western blotting was performed using the following primary antibodies: anti-GFP antibody (AE011, ABclonal Technology, Wuhan, China) and anti-Myc antibody (AE010, ABclonal Technology, Wuhan, China).

Techniques: Activation Assay, Y2H Assay, Transformation Assay, Plasmid Preparation, Bimolecular Fluorescence Complementation Assay, Marker, Co-Immunoprecipitation Assay, Immunoprecipitation, Expressing, Binding Assay