pik3r1  (Bioss)

Journal: European Burn Journal

Article Title: The Role of Angiopoietin-2 in Post-Burn Pneumonia

doi: 10.3390/ebj7010001

Figure Lengend Snippet: Elevated plasma Angiopoietin-2 and downstream pulmonary mediators in a murine model of burn injury. Heparinized plasma was collected one day after burn or sham injury and evaluated for Ang-2 via ELISA. Dark blue dots represent mice that received sham injury, red dots represent mice that received burn injury. ( A ) Plasma Ang-2 was significantly increased in burn mice compared to sham. ( B ) Representative IHC of Ang-2 in sham and burn lungs. ( C – H ) Lung tissue was flash frozen one day after burn injury and then isolated for RNA, and RT-qPCR was performed with GAPDH as housekeeping control. ( C ) RT-qPCR expression of Tek, the transcript for the TIE2 receptor, was significantly elevated in burn lungs compared to sham ( p < 0.01). ( D ) Tek expression positively correlated with plasma Ang-2 levels (simple-linear regression, R 2 = 0.5655, p < 0.01). ( E ) Tnip2, the transcript of ABIN2, and ( F ) Pik3r1, a subunit of PI3K, were both significantly upregulated in the lungs following burn injury. ( G ) ICAM-1 and ( H ) VCAM-1 were also both significantly upregulated in the lungs following burn injury. For two-group analysis, significance was analyzed by unpaired t -test, n = 6–8 animals per group, ** p < 0.01, **** p < 0.0001. ( I – L ) Lung expression of Tnip2, Pik3r1, ICAM-1, and VCAM-1 all positively correlated with plasma Ang-2 levels (simple-linear regression, p < 0.001).

Article Snippet: Pik3r1 , Mm01282781_m1.

Techniques: Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Isolation, Quantitative RT-PCR, Control, Expressing

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A Cell proliferation-related genes in liver tissue of mice exposed to MC-LR in RNA-seq data analysis. B Heatmap of RNA expression of the cell proliferation-related genes. C RT-PCR measurements of the related genes in liver tissue from MC-LR exposed mice. ALKBH5 knockdown reduced PIK3R1 mRNA ( D ) and protein ( E , F ) expression. Overexpression of ALKBH5 increased PIK3R1 mRNA ( G ) and protein ( H , I ) expression. MC-LR exposure resulted in the downregulation of PIK3R1 mRNA ( J ) and protein ( K , L ) expression, which was rescued by overexpression of ALKBH5. M Representative images of immunohistochemistry (IHC) analysis of ALKBH5 and PIK3R1 in liver tissue from MC-LR exposed mice. N IHC staining scores of ALKBH5 and PIK3R1 in liver tissue of mice exposed to MC-LR. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: The detection antibodies were as follows: ALKBH5 antibody (Proteintech, 16837-1-AP); PIK3R1 antibody (Proteintech, 30092-1-AP); MC-LR antibody(Alexis Corporation); β-actin antibody(Cell Signalling Technology, 4967); HK1 antibody (Proteintech, 15656-1-AP); HK2 antibody (Proteintech, 22029-1-AP); PKM antibody (Proteintech, 25659-1-AP); LDHA antibody (Proteintech, 21799-1-AP); ETFDH antibody (Proteintech, 11109-1-AP);NDUFAF4 antibody (Proteintech, 26003-1-AP); Microcystin-LR monoclonal antibody (Enzo Life Sciences, ALX-804-320-C200).

Techniques: RNA Sequencing, RNA Expression, Reverse Transcription Polymerase Chain Reaction, Knockdown, Expressing, Over Expression, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A PIK3R1 mRNA m 6 A methylation was increased upon knockdown of ALKBH5. B PIK3R1 mRNA m 6 A methylation was increased upon MC-LR exposure, which could be rescued by overexpressing ALKBH5. C YTHDF3 knockdown increased PIK3R1 mRNA expression. D YTHDF3 knockdown can reverse the change in PIK3R1 mRNA expression induced by ALKBH5 knockdown. E The half-life of PIK3R1 mRNA was shortened by ALKBH5 knockdown in THLE-3 cells. F The half-life of PIK3R1 mRNA was shortened upon MC-LR exposure, which could be rescued by overexpression of ALKBH5. G Effects of overexpression of ALKBH5 or simultaneous knockdown of PIK3R1 on cell proliferation inhibition induced by MC-LR exposure. Intracellular ATP levels ( H ), extracellular lactate production ( I ) and glucose uptake ( J ) were measured in PIK3R1 knockdown cells compared to control cells. Effects of ALKBH5 overexpression or PIK3R1 knockdown on intracellular ATP levels ( K ) and extracellular lactate production ( L ) in cells exposed to MC-LR. In THLE-3 ( M ) and THLE-2 ( N ) cells, ALKBH5 knockdown reduced the activity of PIK3R1’s WT. The A1557C mutation of PIK3R1 resulted in increased activity and was not affected by ALKBH5 knockdown. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: The detection antibodies were as follows: ALKBH5 antibody (Proteintech, 16837-1-AP); PIK3R1 antibody (Proteintech, 30092-1-AP); MC-LR antibody(Alexis Corporation); β-actin antibody(Cell Signalling Technology, 4967); HK1 antibody (Proteintech, 15656-1-AP); HK2 antibody (Proteintech, 22029-1-AP); PKM antibody (Proteintech, 25659-1-AP); LDHA antibody (Proteintech, 21799-1-AP); ETFDH antibody (Proteintech, 11109-1-AP);NDUFAF4 antibody (Proteintech, 26003-1-AP); Microcystin-LR monoclonal antibody (Enzo Life Sciences, ALX-804-320-C200).

Techniques: Methylation, Knockdown, Expressing, Over Expression, Inhibition, Control, Activity Assay, Mutagenesis

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A RNA-seq data analysis of glycolytic pathway enzyme genes in liver tissue from MC-LR exposed mice. B Heatmap of RNA expression of the glycolytic pathway enzyme genes. C , D The effect of ALKBH5 knockdown on the expression of PIK3R1 and glycolytic pathway enzymes in THLE-3 cells, as detected by Western blotting and quantified. E , F The effect of MC-LR exposure on the expression of MC-LR, ALKBH5, PIK3R1 and glycolytic pathway enzymes in mouse liver tissues, as detected by Western blotting and quantified. G , H The effect of overexpression of ALKBH5 on the expression levels of MC-LR, ALKBH5, PIK3R1 and glycolytic pathway enzymes proteins in THLE-3 cells treated with MC-LR, as detected by Western blotting and quantified. I m 6 A modifications of HK1, HK2, PKM and LDHA were not affected by MC-LR exposure and ALKBH5 overexpression. J , K The effect of PIK3R1 knockdown on the expression of glycolytic pathway enzymes in THLE-3 cells, as detected by Western blotting and quantified. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: The detection antibodies were as follows: ALKBH5 antibody (Proteintech, 16837-1-AP); PIK3R1 antibody (Proteintech, 30092-1-AP); MC-LR antibody(Alexis Corporation); β-actin antibody(Cell Signalling Technology, 4967); HK1 antibody (Proteintech, 15656-1-AP); HK2 antibody (Proteintech, 22029-1-AP); PKM antibody (Proteintech, 25659-1-AP); LDHA antibody (Proteintech, 21799-1-AP); ETFDH antibody (Proteintech, 11109-1-AP);NDUFAF4 antibody (Proteintech, 26003-1-AP); Microcystin-LR monoclonal antibody (Enzo Life Sciences, ALX-804-320-C200).

Techniques: RNA Sequencing, RNA Expression, Knockdown, Expressing, Western Blot, Over Expression

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A , B The effect of PIK3R1 knockdown on the expression of ETFDH, ETFA and NDUFAF4 in THLE-3 cells, as detected by Western blotting and quantified. C , D The effect of overexpression of ALKBH5 on the expression levels of ETFDH, ETFA and NDUFAF4 proteins in THLE-3 cells treated with MC-LR,as detected by Western blotting and quantified. E – G ETFDH, ETFA and NDUFAF4 m 6 A modifications were increased upon knockdown of ALKBH5. H – J ETFDH, ETFA and NDUFAF4 RNA m 6 A modifications was increased upon MC-LR exposure, which could be rescued by overexpressing ALKBH5. K YTHDF3 knockdown increased ETFDH, ETFA and NDUFAF4 mRNA expression. L YTHDF3 knockdown can reverse the change in ETFDH, ETFA and NDUFAF4 mRNA expression induced by ALKBH5 knockdown. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: The detection antibodies were as follows: ALKBH5 antibody (Proteintech, 16837-1-AP); PIK3R1 antibody (Proteintech, 30092-1-AP); MC-LR antibody(Alexis Corporation); β-actin antibody(Cell Signalling Technology, 4967); HK1 antibody (Proteintech, 15656-1-AP); HK2 antibody (Proteintech, 22029-1-AP); PKM antibody (Proteintech, 25659-1-AP); LDHA antibody (Proteintech, 21799-1-AP); ETFDH antibody (Proteintech, 11109-1-AP);NDUFAF4 antibody (Proteintech, 26003-1-AP); Microcystin-LR monoclonal antibody (Enzo Life Sciences, ALX-804-320-C200).

Techniques: Knockdown, Expressing, Western Blot, Over Expression

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A Cell proliferation-related genes in liver tissue of mice exposed to MC-LR in RNA-seq data analysis. B Heatmap of RNA expression of the cell proliferation-related genes. C RT-PCR measurements of the related genes in liver tissue from MC-LR exposed mice. ALKBH5 knockdown reduced PIK3R1 mRNA ( D ) and protein ( E , F ) expression. Overexpression of ALKBH5 increased PIK3R1 mRNA ( G ) and protein ( H , I ) expression. MC-LR exposure resulted in the downregulation of PIK3R1 mRNA ( J ) and protein ( K , L ) expression, which was rescued by overexpression of ALKBH5. M Representative images of immunohistochemistry (IHC) analysis of ALKBH5 and PIK3R1 in liver tissue from MC-LR exposed mice. N IHC staining scores of ALKBH5 and PIK3R1 in liver tissue of mice exposed to MC-LR. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001.

Article Snippet: After deparaffinization and antigen retrieval, the array was incubated with anti-ALKBH5 antibody (dilution 1:200, Proteintech, 16837-1-AP) or anti-PIK3R1 antibody (dilution 1:200, Proteintech, 60225-1-Ig) at 4 °C overnight.

Techniques: RNA Sequencing, RNA Expression, Reverse Transcription Polymerase Chain Reaction, Knockdown, Expressing, Over Expression, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A PIK3R1 mRNA m 6 A methylation was increased upon knockdown of ALKBH5. B PIK3R1 mRNA m 6 A methylation was increased upon MC-LR exposure, which could be rescued by overexpressing ALKBH5. C YTHDF3 knockdown increased PIK3R1 mRNA expression. D YTHDF3 knockdown can reverse the change in PIK3R1 mRNA expression induced by ALKBH5 knockdown. E The half-life of PIK3R1 mRNA was shortened by ALKBH5 knockdown in THLE-3 cells. F The half-life of PIK3R1 mRNA was shortened upon MC-LR exposure, which could be rescued by overexpression of ALKBH5. G Effects of overexpression of ALKBH5 or simultaneous knockdown of PIK3R1 on cell proliferation inhibition induced by MC-LR exposure. Intracellular ATP levels ( H ), extracellular lactate production ( I ) and glucose uptake ( J ) were measured in PIK3R1 knockdown cells compared to control cells. Effects of ALKBH5 overexpression or PIK3R1 knockdown on intracellular ATP levels ( K ) and extracellular lactate production ( L ) in cells exposed to MC-LR. In THLE-3 ( M ) and THLE-2 ( N ) cells, ALKBH5 knockdown reduced the activity of PIK3R1’s WT. The A1557C mutation of PIK3R1 resulted in increased activity and was not affected by ALKBH5 knockdown. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: After deparaffinization and antigen retrieval, the array was incubated with anti-ALKBH5 antibody (dilution 1:200, Proteintech, 16837-1-AP) or anti-PIK3R1 antibody (dilution 1:200, Proteintech, 60225-1-Ig) at 4 °C overnight.

Techniques: Methylation, Knockdown, Expressing, Over Expression, Inhibition, Control, Activity Assay, Mutagenesis

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A RNA-seq data analysis of glycolytic pathway enzyme genes in liver tissue from MC-LR exposed mice. B Heatmap of RNA expression of the glycolytic pathway enzyme genes. C , D The effect of ALKBH5 knockdown on the expression of PIK3R1 and glycolytic pathway enzymes in THLE-3 cells, as detected by Western blotting and quantified. E , F The effect of MC-LR exposure on the expression of MC-LR, ALKBH5, PIK3R1 and glycolytic pathway enzymes in mouse liver tissues, as detected by Western blotting and quantified. G , H The effect of overexpression of ALKBH5 on the expression levels of MC-LR, ALKBH5, PIK3R1 and glycolytic pathway enzymes proteins in THLE-3 cells treated with MC-LR, as detected by Western blotting and quantified. I m 6 A modifications of HK1, HK2, PKM and LDHA were not affected by MC-LR exposure and ALKBH5 overexpression. J , K The effect of PIK3R1 knockdown on the expression of glycolytic pathway enzymes in THLE-3 cells, as detected by Western blotting and quantified. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: After deparaffinization and antigen retrieval, the array was incubated with anti-ALKBH5 antibody (dilution 1:200, Proteintech, 16837-1-AP) or anti-PIK3R1 antibody (dilution 1:200, Proteintech, 60225-1-Ig) at 4 °C overnight.

Techniques: RNA Sequencing, RNA Expression, Knockdown, Expressing, Western Blot, Over Expression

Journal: Cell Death & Disease

Article Title: The dual mechanism of m 6 A demethylase ALKBH5 in regulating energy metabolism during exposure to MC-LR

doi: 10.1038/s41419-025-07791-x

Figure Lengend Snippet: A , B The effect of PIK3R1 knockdown on the expression of ETFDH, ETFA and NDUFAF4 in THLE-3 cells, as detected by Western blotting and quantified. C , D The effect of overexpression of ALKBH5 on the expression levels of ETFDH, ETFA and NDUFAF4 proteins in THLE-3 cells treated with MC-LR,as detected by Western blotting and quantified. E – G ETFDH, ETFA and NDUFAF4 m 6 A modifications were increased upon knockdown of ALKBH5. H – J ETFDH, ETFA and NDUFAF4 RNA m 6 A modifications was increased upon MC-LR exposure, which could be rescued by overexpressing ALKBH5. K YTHDF3 knockdown increased ETFDH, ETFA and NDUFAF4 mRNA expression. L YTHDF3 knockdown can reverse the change in ETFDH, ETFA and NDUFAF4 mRNA expression induced by ALKBH5 knockdown. Data are means ± SD from three independent experiments. * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001.

Article Snippet: After deparaffinization and antigen retrieval, the array was incubated with anti-ALKBH5 antibody (dilution 1:200, Proteintech, 16837-1-AP) or anti-PIK3R1 antibody (dilution 1:200, Proteintech, 60225-1-Ig) at 4 °C overnight.

Techniques: Knockdown, Expressing, Western Blot, Over Expression