Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Immunofluorescence microscopy: ( a ) expression of p53 (green fluorescence) in the cerebral cortex of the control group, which was injected with saline, and experimental groups of animals that were injected with Na 2 S and AOAA, 24 h and 7 days after TBI. Scale bar 20 µm. ( b ) Expression of p53 (green fluorescence) in the cerebral cortex of the control group, which was injected with saline, and experimental groups of animals that were injected with Na 2 S and AOAA, 7 days after TBI. Scale bar 25 µm. ( c ) p53 expression (green fluorescence) in a brain section from an animal injected with AOAA, 7 days after TBI. The white square is the area for analysis of p53 expression. White arrow is the location of the injury. Scale bar 500 µm. ( d ) Coefficient M1 of colocalization of p53 and neuronal nuclear marker NeuN in the contralateral and ipsilateral cortex of the control and experimental groups 24 h and 7 days after TBI. ( e ) Dependence of the average intensity of p53 fluorescence in neurons of the contralateral and ipsilateral cortex of the control and experimental groups 24 h and 7 days after TBI. Contra, contralateral cortex; Ipsi, ipsilateral cortex. NeuN, marker of neuron nuclei (red fluorescence); NeuN+p53 and Hoechst+p53, overlap. Hoechst—Hoechst 33342 fluorescence (blue fluorescence), which visualizes the nuclei of all cells, neurons and glia. One-way ANOVA. M ± SEM. n = 6. ** p < 0.01, *** p < 0.001—ipsilateral cortex relative to contralateral cortex of one animal; # p < 0.05, ## p < 0.01—ipsilateral cortex of the experimental group versus ipsilateral cortex of the control group within one time period after injury.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Immunofluorescence, Microscopy, Expressing, Fluorescence, Control, Injection, Saline, Marker

Journal: International Journal of Molecular Sciences

Article Title: The Role of Hydrogen Sulfide in the Localization and Expression of p53 and Cell Death in the Nervous Tissue in Traumatic Brain Injury and Axotomy

doi: 10.3390/ijms242115708

Figure Lengend Snippet: Immunofluorescence microscopy, TUNEL method. ( a ) Cortical sections stained with TUNEL, control group injected with saline, and experimental groups injected with Na 2 S and AOAA, 24 h and 7 days after TBI, as well as sections stained with TUNEL in the presence of nuclease benzonase (positive control) and in the absence of TdT (negative control). Scale bar 20 µm. ( b ) Cortical sections stained with TUNEL, control group injected with saline, and experimental groups injected with Na 2 S and AOAA, 7 days after TBI. Scale bar 25 µm. ( c ) Cortical section stained with TUNEL, experimental group injected with AOAA, 7 days after TBI. The white square is the analysis area for TUNEL-positive cells. White arrow is the location of the injury. Scale bar 500 µm. ( d ) M1 colocalization coefficient of TUNEL and Hoechst cell nuclear marker in the contralateral and ipsilateral cortex of the control and experimental groups 24 h and 7 days after TBI. ( e ) The number of TUNEL-positive neurons in the contralateral and ipsilateral cortex of the control and experimental groups after 24 h and 7 days of TBI. ( f ) The number of TUNEL-positive glial cells in the contralateral and ipsilateral cortex of the control and experimental groups after 24 h and 7 days of TBI. Scale bar 20 µm. Contra, contralateral cortex; Ipsi, ipsilateral cortex. TUNEL, nuclear marker of cell apoptosis (green fluorescence), NeuN, marker of neuronal nuclei or GFAP marker of astrocytes (red fluorescence); TUNEL+NeuN or TUNEL+GFAP and TUNEL+Hoechst—overlap. Hoechst—Hoechst 33342 fluorescence (blue fluorescence), which visualizes the nuclei of all cells, neurons and glia. One-way ANOVA. M ± SEM. n = 6. * p < 0.05, ** p < 0.01, *** p < 0.001—ipsilateral cortex relative to contralateral cortex of one animal; # p < 0.05, ## p < 0.01—ipsilateral cortex of the experimental group versus ipsilateral cortex of the control group within one time period after injury; and & p < 0.05—ipsilateral cortex of the control and experimental groups versus ipsilateral cortex of the same groups at different time intervals after TBI.

Article Snippet: Sections are then incubated with rabbit primary anti-p53 antibody (1:100; PAA928Mu01, Cloud-Clone Corp, Wuhan, China) and mouse anti-neuN protein antibody (1:1000; FNab10266, FineTest, Wuhan, China) or astrocyte marker GFAP (1:1000; SAB4200571, Sigma Aldrich) for two days at 4 °C.

Techniques: Immunofluorescence, Microscopy, TUNEL Assay, Staining, Control, Injection, Saline, Positive Control, Negative Control, Marker, Fluorescence