x ray irradiator rs 2000  (Rad Source Technologies)

Journal: Inflammation

Article Title: Proline-Mediated Inhibition of ATPIF1-mTOR Signaling Alleviates Radiation-Induced Macrophage Polarization and Colon Inflammation

doi: 10.1007/s10753-025-02404-3

Figure Lengend Snippet: Radiation Induces Macrophage M1 Polarization and Enhances Pro-Inflammatory Cytokine Secretion. ( A ) BMDMs were extracted from the femurs and tibias of C57BL/6 mice and subjected to a 4 Gy dose of X-ray radiation. The percentage of F4/80+/CD86 + macrophages was assessed using FACS analysis. The graph on the right illustrates the representative data showing the proportion of the M1 phenotype. ( B ) RAW264.7 cells were exposed to a 4 Gy dose of X-ray radiation. The percentage of CD86 + cells was determined using FACS analysis. This figure represents one of three independent experiments. ( C ) Western blot analysis showing iNOS protein expression in BMDMs following radiation treatment. β-tubulin was used as a loading control. ( D ) Western blot analysis showing iNOS protein expression in RAW264.7 cells after radiation treatment, with β-tubulin serving as the loading control. ( E ) BMDMs were exposed to a 4 Gy dose of X-ray radiation. The levels of ROS are displayed. ( F ) RAW264.7 were exposed to a 4 Gy dose of X-ray radiation. The levels of ROS are displayed. ( G ) BMDMs were exposed to a 4 Gy dose of X-ray radiation. The levels of mitochondrial ROS are shown. ( H ) RAW264.7 were exposed to a 4 Gy dose of X-ray radiation. The levels of mitochondrial ROS are shown. ( I ) BMDMs were exposed to a 4 Gy dose of X-ray radiation. Shown is the count of Dextran. ( J ) RAW264.7 were exposed to a 4 Gy dose of X-ray radiation. Shown is the count of Dextran. ( K ) BMDMs were exposed to a 4 Gy dose of X-ray radiation. The supernatants from the cell cultures were analyzed for cytokine levels, including TNF-α, IL-1β, and IL-6, using a Cytometric Beads Array. ( L ) RAW264.7 were exposed to a 4 Gy dose of X-ray radiation. Cytokine levels, including TNF-α, IL-1β, and IL-6, were measured in the supernatants from the cell cultures using a Cytometric Beads Array. Error bars are SEM of biological replicates and ** p < 0.01; *** p < 0.001

Article Snippet: Radiation Protocol: The cells were irradiated with 4 Gy of X-rays using an X-RAY irradiator (RAD SOURCE RS 2000 X, USA).

Techniques: Western Blot, Expressing, Control

Journal: Inflammation

Article Title: Proline-Mediated Inhibition of ATPIF1-mTOR Signaling Alleviates Radiation-Induced Macrophage Polarization and Colon Inflammation

doi: 10.1007/s10753-025-02404-3

Figure Lengend Snippet: Proline Supplementation Attenuates Radiation-Induced M1 Polarization and Inflammatory Responses. ( A ) BMDM cells were exposed to a 4 Gy dose of X-ray radiation, either with or without proline treatment. The percentage of F4/80+/CD86 + macrophages was measured using FACS analysis. The graph on the right displays representative data illustrating the proportion of the M1 phenotype. ( B ) RAW264.7 were exposed to a 4 Gy dose of X-ray radiation, either with or without proline treatment. The percentage of CD86 + cells was determined using FACS analysis. This figure represents one of three independent experiments. ( C ) Western blot analysis showing iNOS protein expression in BMDMs following radiation treatment, with or without proline treatment. β-tubulin was used as a loading control. ( D ) Western blot analysis showing iNOS protein expression in RAW264.7 cells following radiation treatment, with or without proline treatment. β-tubulin was used as a loading control. ( E ) BMDMs were exposed to a 4 Gy dose of X-ray radiation, with or without proline treatment. The levels of mitochondrial ROS are presented. ( F ) RAW264.7 were exposed to a 4 Gy dose of X-ray radiation, with or without proline treatment. The levels of mitochondrial ROS are presented. ( G ) BMDMs were exposed to a 4 Gy dose of X-ray radiation, with or without proline treatment. The supernatants from the cell cultures were analyzed for cytokine levels, including TNF-α, IL-1β, and IL-6, using a Cytometric Beads Array. ( H ) RAW264.7 cells received a 4 Gy dose of X-ray radiation, with or without proline treatment. Cytokine levels, including TNF-α, IL-1β, and IL-6, were measured in the cell culture supernatants using a Cytometric Beads Array. Error bars are SEM of biological replicates and * p < 0.05; *** p < 0.001

Article Snippet: Radiation Protocol: The cells were irradiated with 4 Gy of X-rays using an X-RAY irradiator (RAD SOURCE RS 2000 X, USA).

Techniques: Western Blot, Expressing, Control, Cell Culture

Journal: Inflammation

Article Title: Proline-Mediated Inhibition of ATPIF1-mTOR Signaling Alleviates Radiation-Induced Macrophage Polarization and Colon Inflammation

doi: 10.1007/s10753-025-02404-3

Figure Lengend Snippet: Proline Restores Mitochondrial Bioenergetics via Oxidative Phosphorylation Regulation. ( A ) The differences in gene expression between radiation therapy alone and radiation therapy combined with proline were analyzed using RNA sequencing. The displayed results represent the GO term analysis. ( B ) The displayed results represent the KEGG pathway analysis. ( C ) The displayed results represent the enrichment analysis of the cellular pathways associated with the differentially expressed genes. ( D ) The displayed results represent the enrichment analysis of the metabolic pathways related to the differential genes. ( E ) All metabolites between radiation therapy alone and radiation therapy combined with proline were compared using LC/MS-MS. The multiple metabolic pathways involving the different metabolites are displayed. ( F ) RAW264.7 were exposed to a 4 Gy dose of X-ray radiation, with or without proline treatment. The levels of ATP are presented. ( G ) RAW264.7 cells were exposed to a 4 Gy dose of X-ray radiation, with or without proline treatment, and OCR assays were conducted using the Seahorse XF24 analyzer. ( H ) The expression of eight markedly different genes was measured by PCR analysis. Error bars are SEM of biological replicates and * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Radiation Protocol: The cells were irradiated with 4 Gy of X-rays using an X-RAY irradiator (RAD SOURCE RS 2000 X, USA).

Techniques: Phospho-proteomics, Gene Expression, RNA Sequencing, Liquid Chromatography with Mass Spectroscopy, Expressing

Journal: Inflammation

Article Title: Proline-Mediated Inhibition of ATPIF1-mTOR Signaling Alleviates Radiation-Induced Macrophage Polarization and Colon Inflammation

doi: 10.1007/s10753-025-02404-3

Figure Lengend Snippet: Proline Inhibits the Radiation-Activated ATPIF1-mTOR Axis to Attenuate M1 Polarization ( A ) Western blot analysis showing ATPIF1 protein expression in RAW264.7 cells following radiation treatment, with or without proline treatment. β-tubulin was used as a loading control. ( B ) After ATPIF1 knockdown, western blot analysis showing iNOS protein expression in RAW264.7 cells following radiation treatment, with or without proline treatment. β-tubulin was used as a loading control. ( C ) After ATPIF1 knockdown, RAW264.7 were exposed to a 4 Gy dose of X-ray radiation, either with or without proline treatment. The percentage of CD86 + cells was determined using FACS analysis. This figure represents one of three independent experiments. ( D ) Western blot analysis showing mTOR and p-mTOR protein expression in RAW264.7 cells following radiation treatment, with or without proline treatment. β-tubulin was used as a loading control. ( E ) After ATPIF1 knockdown, western blot analysis showing mTOR and p-mTOR protein expression in RAW264.7 cells following radiation treatment, with or without proline treatment. β-tubulin was used as a loading control. ( F ) Western blot analysis showing iNOS protein expression in RAW264.7 cells following radiation, proline and p-mTOR agonist treatment. β-tubulin was used as a loading control. ( G ) Western blot analysis showing iNOS protein expression in RAW264.7 cells following radiation, ATPIF1 knockdown and p-mTOR agonist treatment. β-tubulin was used as a loading control. (H) RAW264.7 cells were exposed to a 4 Gy dose of X-ray radiation, with or without ATPIF1 knockdown, and OCR assays were conducted using the Seahorse XF24 analyzer. Error bars are SEM of biological replicates and * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Radiation Protocol: The cells were irradiated with 4 Gy of X-rays using an X-RAY irradiator (RAD SOURCE RS 2000 X, USA).

Techniques: Western Blot, Expressing, Control, Knockdown