Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: The metabolic remodeling process in natural kidney aging. A Representative images of Oil Red O staining in the 24-month-old and 6-month-old group ( n = 6), scale bar = 50 μm; B , C the downregulated protein levels of FAO markers (i.e., PPARα, ACOX1, and CPT1A) and upregulated protein levels of glycolysis markers (i.e., HK2 and PDK1) by western blot, and their semi-quantitative analyses ( n = 6); D immunostaining for GLIS1 (red) and PPARα (green), with DAPI (blue) counterstaining by IF staining in the 24-month-old and 6-month-old group ( n = 6), scale bar = 50 μm; E lactate levels in the 24-month-old and 6-month-old group ( n = 6); F the downregulated protein levels of PPARα, ACOX1, and CPT1A, and up-regulated protein levels of HK2 and PDK1 in the 24-month-old group by IHC assay and their semi-quantitative analyses ( n = 6), scale bar = 50 μm. The data are expressed as the mean ± SD of three independent experiments. ** P < .01 or *** P < .001 versus the 6-month-old group by Student’s t -test

Article Snippet: Specific primary antibodies targeting GLIS1 (Proteintech 23,138–1-AP), P16INK4A (Thermo Fisher Scientific Cat# MA5-17,142), γ-H2AX (Proteintech 10,856–1-AP), FN (Abcam ab2413), α-SMA (Abcam ab5694), PPARα (Novus NB300-537), ACOX1 (Novus NBP1-80950), CPT1A (Abcam ab234111), HK2 (Sigma SAB2108077), PDK1 (Sigma SAB4502160), METTL3 (Abcam ab195352), and YTHDF1 (Abcam ab252346) were incubated overnight at 4 °C.

Techniques: Staining, Western Blot, Immunostaining

Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: GLIS1 regulated metabolic remodeling from FAO to glycolysis pathway by binding to the PPARα promoter. A Representative images of Oil Red O staining in the control vector and siGLIS1 group ( n = 3), scale bar = 50 μm; B , C the ECAR levels after culturing with glucose followed by oligomycin and 2-DG in the control vector and siGLIS1 group, and its semi-quantitative analysis ( n = 3); D metabolic component analysis in control and siGLIS1 group ( n = 3); E , F downregulated protein levels of GLIS1, PPARα, ACOX1, and CPTA1, and upregulated protein levels of HK2 and PDK1 in siGLIS1 group by western blot, and their semi-quantitative analyses ( n = 3); G – I cellular free fatty acid (FFA), triglyceride (TG) and total cholesterol (TC) levels in HK-2 cells in control and siGLIS1 group ( n = 3); J immunostaining for PPARα, ACOX1, CPT1A, HK2, and PDK1 (green), with DAPI (blue) counterstaining by IF staining ( n = 3), scale bar = 50 μm; K the interaction of PPARα and GLIS1 confirmed by ChIP and PCR assay ( n = 3); L the protein levels of FN and HK2 by western blot in HK-2 cells treated with D-gal in the presence of OE-GLIS1, OE-GLIS1 + CPTA1 inhibitor (etomoxir), OE-GLIS1 + PPARα (MK886), and OE-GLIS1 + siACOX1( n = 3). The data are expressed as the mean ± SD of three independent experiments. ** P < .01 or *** P < .001 versus the vector group ( C , D , F , H , and I ), or IgG group ( K ) by Student’s t -test

Article Snippet: Specific primary antibodies targeting GLIS1 (Proteintech 23,138–1-AP), P16INK4A (Thermo Fisher Scientific Cat# MA5-17,142), γ-H2AX (Proteintech 10,856–1-AP), FN (Abcam ab2413), α-SMA (Abcam ab5694), PPARα (Novus NB300-537), ACOX1 (Novus NBP1-80950), CPT1A (Abcam ab234111), HK2 (Sigma SAB2108077), PDK1 (Sigma SAB4502160), METTL3 (Abcam ab195352), and YTHDF1 (Abcam ab252346) were incubated overnight at 4 °C.

Techniques: Binding Assay, Staining, Control, Plasmid Preparation, Western Blot, Immunostaining

Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: The over-expressed GLIS1 suppressed metabolic remodeling from FAO to glycolysis in the accelerated aging mouse model. A The representative images of mouse kidney tissue in the control, AAV-Vector and AAV-GLIS1 group stained with Oil Red O ( n = 6), scale bar = 50 μm; B , C downregulated protein levels of PPARα, ACOX1, and CPT1A in the accelerated aging mouse model were reversed in the presence of AAV-GLIS1, while upregulated protein levels of HK2 and PKD1 in the accelerated aging mouse model were reduced by introducing AAV-GLIS1 ( n = 6); D lactate levels in the control, AAV-Vector and AAV-GLIS1 group ( n = 6); E the reversed effect of PPARα, ACOX1 and CPT1A levels, as well as HK2 and PDK1 levels in the presence of AAV-GLIS1 by IHC assay, and their semi-quantitative analyses ( n = 6), scale bar = 50 μm; F immunostaining for PPARα, ACOX1, CPTA1, HK2, and PDK1 (green), with DAPI (blue) counterstaining by IF staining in accelerated aging mouse model ( n = 6), scale bar = 50 μm. The data are expressed as the mean ± SD of three independent experiments. ** P < .01 or *** P < .001 versus the AAV-vector group by one-way ANOVA

Article Snippet: Specific primary antibodies targeting GLIS1 (Proteintech 23,138–1-AP), P16INK4A (Thermo Fisher Scientific Cat# MA5-17,142), γ-H2AX (Proteintech 10,856–1-AP), FN (Abcam ab2413), α-SMA (Abcam ab5694), PPARα (Novus NB300-537), ACOX1 (Novus NBP1-80950), CPT1A (Abcam ab234111), HK2 (Sigma SAB2108077), PDK1 (Sigma SAB4502160), METTL3 (Abcam ab195352), and YTHDF1 (Abcam ab252346) were incubated overnight at 4 °C.

Techniques: Control, Plasmid Preparation, Staining, Immunostaining

Journal: BMC Biology

Article Title: N6-methyladenosine regulates metabolic remodeling in kidney aging through transcriptional regulator GLIS1

doi: 10.1186/s12915-024-02100-y

Figure Lengend Snippet: The ablation of METTL3 triggered metabolic remodeling and aggravated cell senescence renal fibrosis. A Protein levels of GLIS1, PPARα, ACOX1, CPT1A, HK2, PDK1, FN, α-SMA, and P16 INK4A by western blot in the vector and siMETTL3 group, and their semi-quantitative analyses ( n = 3); B representative images of Oil Red O staining in the vector and siMETTL3 group ( n = 3), scale bar = 50 μm; C double immunostaining for GLS1 and PPARα, ACOX1, CPT1A, HK2, PDK1, and α-SMA by IF staining ( n = 3) scale bar = 50 μm. The data are expressed as the mean ± SD of three independent experiments. ** P < .01 or *** P < .001 versus the vector group by Student’s t -test

Article Snippet: Specific primary antibodies targeting GLIS1 (Proteintech 23,138–1-AP), P16INK4A (Thermo Fisher Scientific Cat# MA5-17,142), γ-H2AX (Proteintech 10,856–1-AP), FN (Abcam ab2413), α-SMA (Abcam ab5694), PPARα (Novus NB300-537), ACOX1 (Novus NBP1-80950), CPT1A (Abcam ab234111), HK2 (Sigma SAB2108077), PDK1 (Sigma SAB4502160), METTL3 (Abcam ab195352), and YTHDF1 (Abcam ab252346) were incubated overnight at 4 °C.

Techniques: Western Blot, Plasmid Preparation, Staining, Double Immunostaining

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: KEGG pathway analysis of differentially expressed genes associated with NSCLC

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques: Starch

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: The expression levels of mRNA COL12A1 (G) , COL1A2 (D) , COL4A1 (E) , SPARC (K) , COL5A2 (F) and COL1A1 (C) was increased, while ACAA1 (A) , ACOX1 (B) , CPT2 (H) , SQRDL (L) , HMGCS2 (J) , ETHE1 (I) and TST (M) were decreased. * P < 0.05.

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques: Expressing

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: The expression levels of protein COL12A1 (G) , COL1A2 (D) , COL4A1 (E) , SPARC (K) , COL5A2 (F) and COL1A1 (C) was increased, while ACAA1 (A) , ACOX1 (B) , CPT2 (H) , SQRDL (L) , HMGCS2 (J) , ETHE1 (I) and TST (M) were decreased. * P < 0.05.

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques: Expressing

Journal: Oncotarget

Article Title: hsa-miR-29c-3p regulates biological function of colorectal cancer by targeting SPARC

doi: 10.18632/oncotarget.22356

Figure Lengend Snippet: Primer pairs of different genes

Article Snippet: Membranes were performed blockage whit 5% skim milk for 3 h and then incubation with primary antibodies for ACAA1 (ab110289, Abcam, 1:1000) antibody, ACOX1 (ab184032, Abcam, 1:1000) antibody, COL1A1 (PB0980, Boster, 1:500) antibody, COL1A2 (ab208638, Abcam, 1:500) antibody, COL4A1 (PB0126, Boster, 1:500) antibody, COL5A2 (A03869, Boster, 1:1000) antibody, COL12A1 (ab121304, Abcam, 1:1000) antibody, CPT2 (ab181114, Abcam, 1:4000) antibody, ETHE1 (ab174302, Abcam, 1:4000) antibody, HMGCS2 (ab137043, Abcam, 1:4000) antibody, SPARC (ab207743, Abcam, 1:1000) antibody, SQRDL (ab71978, Abcam, 1:500) antibody, TST (ab166625, Abcam, 1:4000) antibody and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, 1:4000) antibody, and subsequent incubation whit horseradish peroxidase (HRP)-coupled goat anti-rabbit (ZDR-5306, Beijing ZSBio, 1:5000) or HRP-coupled goat anti-mouse (W420B, Promega, 1:5000) secondary antibody, respectively.

Techniques: