Journal: Frontiers in Genetics

Article Title: Exploring the prognostic significance of lactate-mitochondria-related genes in prostate cancer

doi: 10.3389/fgene.2024.1515045

Figure Lengend Snippet: MPO inhibits prostate cancer metastasis. (A) The cell number of LNCaP cells was counted at different time points. (B, C) Cell cycle analysis through PI staining was detected by FACS (B) , and the cell cycle distribution was presented (C) . (D, E) LNCaP cells were analyzed for cell migration and invasion. Representative images of crystal violet-stained migrated (D) or invaded (E) cells are presented (scale bar, 100 μL). (F) The Tyr925-phosphorylated level of FAK and total FAK, E-Cadherin, and Snail in LNCaP cells were detected by Western blot analysis. Actin was used as a loading control. All data are presented as the mean ± SD of three independent experiments; ns, not significant.

Article Snippet: Anti- FAK (Cat# 3285), anti-pFAK Tyr925 (Cat# 3284), anti-Snail (Cat# 3879), anti-E-Cadherin (Cat# 3195) and anti-actin (Cat# 4970) antibodies were purchased from Cell Signaling Technology.

Techniques: Cell Cycle Assay, Staining, Migration, Western Blot, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Resolvin E1 is a pro-repair molecule that promotes intestinal epithelial wound healing

doi: 10.1073/pnas.1921335117

Figure Lengend Snippet: RvE1 promotes intestinal epithelial wound repair. (A) Wound areas of scratch-wounded SKCO15 IEC monolayers, incubated with increasing concentrations (10, 50, 100, and 500 nM) of RvE1 were continuously imaged. Percentage wound closure was calculated by comparison of 0 and 24 h postinjury. (B) Wounded primary IECs treated with RvE1 100 nM or vehicle for 24 h. (C) Scratch-wounded intestinal epithelial monolayers were treated with RvE1 (100 nM) or vehicle, and EdU incorporation was determined 24 h postwound ***P < 0.001; mean ± SEM. (D) Immunoblotting was performed on lysates from scratch-wounded SKCO15 monolayers treated with RvE1 (100 nM) or vehicle for different points. Levels of pP70, pmTOR, pCreb, pSRC (416), and pFAK (Y397, Y925) were compared with total P70, mTOR, CREB, Src, FAK, and GAPDH to assess activation. Densitometry values are displayed under the phosphorylated protein blots; values are normalized to total protein and nonwounded cells except for FAK blots, which were normalized to loading control. (E) SKCO15 IECs were incubated with RvE1 (100 nM) or vehicle for 4 h. ROS generation was detected by confocal microscopy, using the fluorescent hydro-Cy3 dye in scratch-wounded monolayers adjacent to the wound edge. Quantification was done calculating the fold change increase in pixel counts of RvE1 treatment compared with vehicle, using ImageJ software. **P < 0.01; ***P < 0.001; mean ± SEM.

Article Snippet: The following antibodies were used: FAK (cat. 610088; BD Biosciences); pFAK (Y861; cat. PS 1008; Calbiochem); pFAK (Tyr397; cat. 3283), pFAK (Tyr925; cat. 3284), Src (cat. 2108), pSrc (Tyr416; cat. 2101), p70 S6 Kinase (cat. 9202), p-p70 S6 Kinase (cat. 9209), mTOR (cat. 2983), pmTOR (cat. 2448), CREB (cat. 9197), and pCREB (cat. 87G3; Cell Signaling Technology); and claudin 4 (cat. 364800; Invitrogen).

Techniques: Incubation, Western Blot, Activation Assay, Confocal Microscopy, Software

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Resolvin E1 is a pro-repair molecule that promotes intestinal epithelial wound healing

doi: 10.1073/pnas.1921335117

Figure Lengend Snippet: RvE1 promotes intestinal epithelial wound repair. (A) Wound areas of scratch-wounded SKCO15 IEC monolayers, incubated with increasing concentrations (10, 50, 100, and 500 nM) of RvE1 were continuously imaged. Percentage wound closure was calculated by comparison of 0 and 24 h postinjury. (B) Wounded primary IECs treated with RvE1 100 nM or vehicle for 24 h. (C) Scratch-wounded intestinal epithelial monolayers were treated with RvE1 (100 nM) or vehicle, and EdU incorporation was determined 24 h postwound ***P < 0.001; mean ± SEM. (D) Immunoblotting was performed on lysates from scratch-wounded SKCO15 monolayers treated with RvE1 (100 nM) or vehicle for different points. Levels of pP70, pmTOR, pCreb, pSRC (416), and pFAK (Y397, Y925) were compared with total P70, mTOR, CREB, Src, FAK, and GAPDH to assess activation. Densitometry values are displayed under the phosphorylated protein blots; values are normalized to total protein and nonwounded cells except for FAK blots, which were normalized to loading control. (E) SKCO15 IECs were incubated with RvE1 (100 nM) or vehicle for 4 h. ROS generation was detected by confocal microscopy, using the fluorescent hydro-Cy3 dye in scratch-wounded monolayers adjacent to the wound edge. Quantification was done calculating the fold change increase in pixel counts of RvE1 treatment compared with vehicle, using ImageJ software. **P < 0.01; ***P < 0.001; mean ± SEM.

Article Snippet: The following antibodies were used: FAK (cat. 610088; BD Biosciences); pFAK (Y861; cat. PS 1008; Calbiochem); pFAK (Tyr397; cat. 3283), pFAK (Tyr925; cat. 3284), Src (cat. 2108), pSrc (Tyr416; cat. 2101), p70 S6 Kinase (cat. 9202), p-p70 S6 Kinase (cat. 9209), mTOR (cat. 2983), pmTOR (cat. 2448), CREB (cat. 9197), and pCREB (cat. 87G3; Cell Signaling Technology); and claudin 4 (cat. 364800; Invitrogen).

Techniques: Incubation, Western Blot, Activation Assay, Confocal Microscopy, Software

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Resolvin E1 is a pro-repair molecule that promotes intestinal epithelial wound healing

doi: 10.1073/pnas.1921335117

Figure Lengend Snippet: RvE1 promotes intestinal epithelial wound repair. (A) Wound areas of scratch-wounded SKCO15 IEC monolayers, incubated with increasing concentrations (10, 50, 100, and 500 nM) of RvE1 were continuously imaged. Percentage wound closure was calculated by comparison of 0 and 24 h postinjury. (B) Wounded primary IECs treated with RvE1 100 nM or vehicle for 24 h. (C) Scratch-wounded intestinal epithelial monolayers were treated with RvE1 (100 nM) or vehicle, and EdU incorporation was determined 24 h postwound ***P < 0.001; mean ± SEM. (D) Immunoblotting was performed on lysates from scratch-wounded SKCO15 monolayers treated with RvE1 (100 nM) or vehicle for different points. Levels of pP70, pmTOR, pCreb, pSRC (416), and pFAK (Y397, Y925) were compared with total P70, mTOR, CREB, Src, FAK, and GAPDH to assess activation. Densitometry values are displayed under the phosphorylated protein blots; values are normalized to total protein and nonwounded cells except for FAK blots, which were normalized to loading control. (E) SKCO15 IECs were incubated with RvE1 (100 nM) or vehicle for 4 h. ROS generation was detected by confocal microscopy, using the fluorescent hydro-Cy3 dye in scratch-wounded monolayers adjacent to the wound edge. Quantification was done calculating the fold change increase in pixel counts of RvE1 treatment compared with vehicle, using ImageJ software. **P < 0.01; ***P < 0.001; mean ± SEM.

Article Snippet: The following antibodies were used: FAK (cat. 610088; BD Biosciences); pFAK (Y861; cat. PS 1008; Calbiochem); pFAK (Tyr397; cat. 3283), pFAK (Tyr925; cat. 3284), Src (cat. 2108), pSrc (Tyr416; cat. 2101), p70 S6 Kinase (cat. 9202), p-p70 S6 Kinase (cat. 9209), mTOR (cat. 2983), pmTOR (cat. 2448), CREB (cat. 9197), and pCREB (cat. 87G3; Cell Signaling Technology); and claudin 4 (cat. 364800; Invitrogen).

Techniques: Incubation, Western Blot, Activation Assay, Confocal Microscopy, Software

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Resolvin E1 is a pro-repair molecule that promotes intestinal epithelial wound healing

doi: 10.1073/pnas.1921335117

Figure Lengend Snippet: RvE1 promotes intestinal epithelial wound repair. (A) Wound areas of scratch-wounded SKCO15 IEC monolayers, incubated with increasing concentrations (10, 50, 100, and 500 nM) of RvE1 were continuously imaged. Percentage wound closure was calculated by comparison of 0 and 24 h postinjury. (B) Wounded primary IECs treated with RvE1 100 nM or vehicle for 24 h. (C) Scratch-wounded intestinal epithelial monolayers were treated with RvE1 (100 nM) or vehicle, and EdU incorporation was determined 24 h postwound ***P < 0.001; mean ± SEM. (D) Immunoblotting was performed on lysates from scratch-wounded SKCO15 monolayers treated with RvE1 (100 nM) or vehicle for different points. Levels of pP70, pmTOR, pCreb, pSRC (416), and pFAK (Y397, Y925) were compared with total P70, mTOR, CREB, Src, FAK, and GAPDH to assess activation. Densitometry values are displayed under the phosphorylated protein blots; values are normalized to total protein and nonwounded cells except for FAK blots, which were normalized to loading control. (E) SKCO15 IECs were incubated with RvE1 (100 nM) or vehicle for 4 h. ROS generation was detected by confocal microscopy, using the fluorescent hydro-Cy3 dye in scratch-wounded monolayers adjacent to the wound edge. Quantification was done calculating the fold change increase in pixel counts of RvE1 treatment compared with vehicle, using ImageJ software. **P < 0.01; ***P < 0.001; mean ± SEM.

Article Snippet: The following antibodies were used: FAK (cat. 610088; BD Biosciences); pFAK (Y861; cat. PS 1008; Calbiochem); pFAK (Tyr397; cat. 3283), pFAK (Tyr925; cat. 3284), Src (cat. 2108), pSrc (Tyr416; cat. 2101), p70 S6 Kinase (cat. 9202), p-p70 S6 Kinase (cat. 9209), mTOR (cat. 2983), pmTOR (cat. 2448), CREB (cat. 9197), and pCREB (cat. 87G3; Cell Signaling Technology); and claudin 4 (cat. 364800; Invitrogen).

Techniques: Incubation, Western Blot, Activation Assay, Confocal Microscopy, Software

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Resolvin E1 is a pro-repair molecule that promotes intestinal epithelial wound healing

doi: 10.1073/pnas.1921335117

Figure Lengend Snippet: RvE1 promotes intestinal epithelial wound repair. (A) Wound areas of scratch-wounded SKCO15 IEC monolayers, incubated with increasing concentrations (10, 50, 100, and 500 nM) of RvE1 were continuously imaged. Percentage wound closure was calculated by comparison of 0 and 24 h postinjury. (B) Wounded primary IECs treated with RvE1 100 nM or vehicle for 24 h. (C) Scratch-wounded intestinal epithelial monolayers were treated with RvE1 (100 nM) or vehicle, and EdU incorporation was determined 24 h postwound ***P < 0.001; mean ± SEM. (D) Immunoblotting was performed on lysates from scratch-wounded SKCO15 monolayers treated with RvE1 (100 nM) or vehicle for different points. Levels of pP70, pmTOR, pCreb, pSRC (416), and pFAK (Y397, Y925) were compared with total P70, mTOR, CREB, Src, FAK, and GAPDH to assess activation. Densitometry values are displayed under the phosphorylated protein blots; values are normalized to total protein and nonwounded cells except for FAK blots, which were normalized to loading control. (E) SKCO15 IECs were incubated with RvE1 (100 nM) or vehicle for 4 h. ROS generation was detected by confocal microscopy, using the fluorescent hydro-Cy3 dye in scratch-wounded monolayers adjacent to the wound edge. Quantification was done calculating the fold change increase in pixel counts of RvE1 treatment compared with vehicle, using ImageJ software. **P < 0.01; ***P < 0.001; mean ± SEM.

Article Snippet: The following antibodies were used: FAK (cat. 610088; BD Biosciences); pFAK (Y861; cat. PS 1008; Calbiochem); pFAK (Tyr397; cat. 3283), pFAK (Tyr925; cat. 3284), Src (cat. 2108), pSrc (Tyr416; cat. 2101), p70 S6 Kinase (cat. 9202), p-p70 S6 Kinase (cat. 9209), mTOR (cat. 2983), pmTOR (cat. 2448), CREB (cat. 9197), and pCREB (cat. 87G3; Cell Signaling Technology); and claudin 4 (cat. 364800; Invitrogen).

Techniques: Incubation, Western Blot, Activation Assay, Confocal Microscopy, Software

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Resolvin E1 is a pro-repair molecule that promotes intestinal epithelial wound healing

doi: 10.1073/pnas.1921335117

Figure Lengend Snippet: RvE1 promotes intestinal epithelial wound repair. (A) Wound areas of scratch-wounded SKCO15 IEC monolayers, incubated with increasing concentrations (10, 50, 100, and 500 nM) of RvE1 were continuously imaged. Percentage wound closure was calculated by comparison of 0 and 24 h postinjury. (B) Wounded primary IECs treated with RvE1 100 nM or vehicle for 24 h. (C) Scratch-wounded intestinal epithelial monolayers were treated with RvE1 (100 nM) or vehicle, and EdU incorporation was determined 24 h postwound ***P < 0.001; mean ± SEM. (D) Immunoblotting was performed on lysates from scratch-wounded SKCO15 monolayers treated with RvE1 (100 nM) or vehicle for different points. Levels of pP70, pmTOR, pCreb, pSRC (416), and pFAK (Y397, Y925) were compared with total P70, mTOR, CREB, Src, FAK, and GAPDH to assess activation. Densitometry values are displayed under the phosphorylated protein blots; values are normalized to total protein and nonwounded cells except for FAK blots, which were normalized to loading control. (E) SKCO15 IECs were incubated with RvE1 (100 nM) or vehicle for 4 h. ROS generation was detected by confocal microscopy, using the fluorescent hydro-Cy3 dye in scratch-wounded monolayers adjacent to the wound edge. Quantification was done calculating the fold change increase in pixel counts of RvE1 treatment compared with vehicle, using ImageJ software. **P < 0.01; ***P < 0.001; mean ± SEM.

Article Snippet: The following antibodies were used: FAK (cat. 610088; BD Biosciences); pFAK (Y861; cat. PS 1008; Calbiochem); pFAK (Tyr397; cat. 3283), pFAK (Tyr925; cat. 3284), Src (cat. 2108), pSrc (Tyr416; cat. 2101), p70 S6 Kinase (cat. 9202), p-p70 S6 Kinase (cat. 9209), mTOR (cat. 2983), pmTOR (cat. 2448), CREB (cat. 9197), and pCREB (cat. 87G3; Cell Signaling Technology); and claudin 4 (cat. 364800; Invitrogen).

Techniques: Incubation, Western Blot, Activation Assay, Confocal Microscopy, Software