Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: FOSL1 promotes pDCs activation by mediating the oxidative stress of pDCs. ( A ) MDA and SOD concentrations in the supernatant of mouse pancreatic tissue homogenate ( n = 5). ( B ) Analysis of H 2 O 2 and GSH levels in mouse pancreatic tissue homogenate supernatant ( n = 5). ( C ) Analysis of ROS content within mouse pDCs by DCFH-DA probe. Statistical analyses were performed using one-way ANOVA, followed by Tukey’s post hoc test. The results shown are representative of data derived from 3 independent experiments and are expressed as mean ± SEM. * p < 0.05.

Article Snippet: The cells were stained with fluorescent probe DCFH-DA (HY-D0940, MedChemExpress) for 30 min and washed with PBS.

Techniques: Activation Assay, Derivative Assay

Journal: Inflammation

Article Title: Bifidobacterium Infantis Promotes FOSL1-Mediated Transcriptional Repression of VDR to Accelerate Autoimmune Pancreatitis Progression Through Activation of Oxidative Stress

doi: 10.1007/s10753-025-02368-4

Figure Lengend Snippet: Knockdown of VDR expedites oxidative stress in pDCs in the presence of FOSL1 knockdown. ( A ) The content of IFN-α and IL-33 secreted by pDCs transduced with sh-FOSL1 + sh-Scramble or sh-VDR was examined using ELISA. ( B ) VDR expression in isolated pDCs (marked by pDCA-1) was observed using dual-labeling immunofluorescence. ( C ) Activated pDCs (Siglec-H + ) in total pDCs were analyzed using flow cytometry. ( D ) Analysis of ROS content within mouse pDCs by DCFH-DA probe. Statistical analyses were performed using unpaired t-tests. The results shown are representative of data derived from 3 independent experiments and are expressed as mean ± SEM. * p < 0.05.

Article Snippet: The cells were stained with fluorescent probe DCFH-DA (HY-D0940, MedChemExpress) for 30 min and washed with PBS.

Techniques: Knockdown, Transduction, Enzyme-linked Immunosorbent Assay, Expressing, Isolation, Labeling, Immunofluorescence, Flow Cytometry, Derivative Assay

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: The protective effects of antioxidants against endogenous and exogenous oxidative stress on bull sperm

doi: 10.1007/s11626-024-00944-w

Figure Lengend Snippet: Impact of various antioxidants on fresh bull sperm motility and reactive oxygen species (ROS) levels when exposed to different oxidative stress triggers. ( A ) Experimental design to examine the impact of various antioxidants on sperm motility and levels of reactive oxygen species (ROS) under the two oxidative stress inducers. ( B and C ) Changes in sperm viability and motility under H 2 O 2 ( B ) or antimycin (C). ( D - G ) Effect of antioxidants on the percentages of DCFH-DA and mtSOX deep Red positive sperm under H 2 O 2 or antimycin. ( H - J ) Measurement of mitochondrial activity using the JC-1 kit. ( H ) Representative flow cytometric pattern of sperm that was not treated with any drug (control) stained with JC-1. Effect of antioxidants on the percentages of high mitochondrial membrane potential (hMMP) under the treatment of H 2 O 2 ( I ) and antimycin ( J ). H 2 O 2 or antimycin groups were used as controls for statistical analysis ( p < 0.05).

Article Snippet: Photooxidation-resistant dichloro-dihydro-fluorescein diacetate (DCFH-DA) is a total ROS (H 2 O 2 , superoxide anion, and hydroxyl radical) probe (R253; DOJINDO LABOLATORIES Co., Ltd., Kumamoto, Japan).

Techniques: Activity Assay, Control, Staining, Membrane

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: The protective effects of antioxidants against endogenous and exogenous oxidative stress on bull sperm

doi: 10.1007/s11626-024-00944-w

Figure Lengend Snippet: The effect of various antioxidants on frozen-thawed bull sperm motility and ROS levels. ( A ) Experimental design to examine the impact of PQQ, vitamin C, and ergothioneine on frozen-thawed bull sperm motility and ROS levels. Various antioxidants were included in all thawing, washing, and incubation processes using HTF medium as the basic medium. ( B and C ) Effect of antioxidants on the percentages of DCFH-DA ( B ) and mtSOX deep Red ( C ) positive sperm. ( D ) Changes in frozen-thawed sperm viability and kinetics patterns with antioxidants. Asterisks represent significant differences from the control (Ctrl; *: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: Photooxidation-resistant dichloro-dihydro-fluorescein diacetate (DCFH-DA) is a total ROS (H 2 O 2 , superoxide anion, and hydroxyl radical) probe (R253; DOJINDO LABOLATORIES Co., Ltd., Kumamoto, Japan).

Techniques: Incubation, Control

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: The protective effects of antioxidants against endogenous and exogenous oxidative stress on bull sperm

doi: 10.1007/s11626-024-00944-w

Figure Lengend Snippet: Significant decrease in viability and an increase in ROS levels of fresh bull sperm during the incubation period. ( A ) Tracks of sperm incubated for 0 and 4 h were determined using the CASA system. Red arrows indicate the aggregated sperm, which is the cause of overestimating CASA parameters because it lowers the number of dead sperm. ( B ) Changes in fresh sperm motility single counts implying sperm viability and in kinetics patterns with incubation time. ( C and D ) Time-dependent changes in total ROS in the cytoplasm or mitochondrial superoxide level of fresh bull sperm detected using DCFH-DA or mtSOX deep Red, respectively. Flow cytometric patterns and percentage of positive sperm were shown. Different letters represent significantly different groups ( p < 0.05).

Article Snippet: Photooxidation-resistant dichloro-dihydro-fluorescein diacetate (DCFH-DA) is a total ROS (H 2 O 2 , superoxide anion, and hydroxyl radical) probe (R253; DOJINDO LABOLATORIES Co., Ltd., Kumamoto, Japan).

Techniques: Incubation

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: The protective effects of antioxidants against endogenous and exogenous oxidative stress on bull sperm

doi: 10.1007/s11626-024-00944-w

Figure Lengend Snippet: The impact of oxidative stress inducers, H 2 O 2 and antimycin, on fresh bull sperm motility and ROS levels. ( A ) Experimental design to investigate the effect of the oxidative stress inducers on fresh bull sperm. ( B ) Representative images of the sperm fluorescence signal with DCFH-DA and mtSOX deep Red. Left : DCFH-DA fluorescence image of sperm treated with H 2 O 2 . Right : mtSOX deep Red fluorescence image of sperm treated with antimycin. ( C ) Flow cytometric patterns of DCFH-DA. ( D ) Percentages of DCFH-DA positive sperm. ( E ) Sperm kinetics changed by H 2 O 2 . ( F ) Flow cytometric patterns of mtSOX deep Red. ( G ) Percentages of mtSOX deep Red positive sperm. ( H ) Sperm kinetics changed by antimycin. Different letters represent significantly different groups ( p < 0.05).

Article Snippet: Photooxidation-resistant dichloro-dihydro-fluorescein diacetate (DCFH-DA) is a total ROS (H 2 O 2 , superoxide anion, and hydroxyl radical) probe (R253; DOJINDO LABOLATORIES Co., Ltd., Kumamoto, Japan).

Techniques: Fluorescence

Journal: In Vitro Cellular & Developmental Biology. Animal

Article Title: The protective effects of antioxidants against endogenous and exogenous oxidative stress on bull sperm

doi: 10.1007/s11626-024-00944-w

Figure Lengend Snippet: Impact of antioxidants on total ROS and mitochondrial ROS levels in fresh bull sperm under oxidative stress inducers. ( A ) Experimental design to investigate the effect of three antioxidants on induced reactive oxygen species in fresh bull sperm. ( B ) Flow cytometric patterns of H 2 O 2 and antioxidant-treated sperm stained with DCFH-DA. ( C ) Percentages of DCFH-DA positive sperm. ( D ) Flow cytometric patterns of antimycin and antioxidant-treated sperm stained with mtSOX deep Red. ( E ) Percentages of mtSOX deep Red positive sperm. Different letters represent significantly different groups ( p < 0.05).

Article Snippet: Photooxidation-resistant dichloro-dihydro-fluorescein diacetate (DCFH-DA) is a total ROS (H 2 O 2 , superoxide anion, and hydroxyl radical) probe (R253; DOJINDO LABOLATORIES Co., Ltd., Kumamoto, Japan).

Techniques: Staining