Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 3 Exosome and exosome-derived circHIF1α inhibits PK-15 cells proliferation. A–B The relative expression of circHIF1α in the precipitation or super natant of PIEC/PK-15 cells after PIEC cells treated with G. parasuis. C The relative expression of circHIF1α in PK-15 cells treated with exosomes from over expressed circHIF1α PIEC cells. D–G The proliferation of PK-15 cells treated with exosomes from PIEC cells after circHIF1α overexpression or knockdown was evaluated by CCK-8 (D and F) and EdU assay (E and G). Scale bar, 75 μm. H–I. The proliferation of PK-15 cells transfected with circHIF1α siRNA or overexpression vector was evaluated by CCK-8 (H) and EdU assay (I). Scale bar, 100 μm. J Mass spectrometric data of interaction between circHIF1α and ptotein showed the important correlated biological process. K The pathway result of mass spectrometry after RNA pulldown assay

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Derivative Assay, Expressing, Over Expression, Knockdown, CCK-8 Assay, EdU Assay, Transfection, Plasmid Preparation, Mass Spectrometry

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 4 Exosome and exosome-derived circHIF1α promotes PK-15 cells DNA damage, and mediates the G1/S phase. A–B The DNA damage level of PK-15 cells treated with exosomes from PIEC cells after circHIF1α overexpression or knockdown was detected by γ-H2AX immunofluorescence. Scale bar, 75 μm. C Cell cycle analysis was executed by flow cytometry when PK-15 cells were treated with exosomes from PIEC cells after circHIF1α knockdown. D Western blot showing the levels of DNA damage-related proteins, including γ-H2AX, NPM1, p53, and p-p53 in PK-15 cells transfected with ov-circHIF1α or ov-pLC5. E Cell cycle analysis was executed by flow cytometry after circHIF1α overexpression for 24 h. F The expression level of cell cycle-related proteins was detected in PK-15 cells after circHIF1α overexpression for 24 h by Western blot. Data are represented as mean ± SD. NS, not signifcant, *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Derivative Assay, Over Expression, Knockdown, Immunofluorescence, Cell Cycle Assay, Flow Cytometry, Western Blot, Transfection, Expressing

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 5 CircHIF1α reduces bacterial adhesion and invasion for PK-15 cell. A–C The quantity of PK-15 cells infected with G. parasuis (A)/S. aureus (B)/SS2 (C) was identified by the viable counting method. D–F PK-15 cells infected with G. parasuis(D) /S. aureus (E)/SS2 (F) was identified by IF. Scale bar, 25 μm. G–I The quantity of PK-15 cells infected with G. parasuis (G) /S. aureus (H) /SS2 (I) was identified after PK-15 cells were treated with 250nM NSC 80,467 by bacteria adhesion and invasion assays. J–L. The quantity of PK-15 cells infected with G. parasuis (J), S. aureus (K), and SS2 (L) was identified after PK-15 cells were treated with 2mM Thymidine by bacteria adhesion and invasion assays. Data are represented as mean ± SD. NS, not signifcant, *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Infection, Bacteria

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 8 Exosomal circHIF1α resists bacterial infection in vivo. A Left, Bioluminescent image showed localization of G. parasuis-exosome (20 mg/kg) or Mock-exosome (20 mg/kg) in mice after injecting through the tail vein at different times. Right, the fluorescence intensity of exosomes in vivo. n = 6 mice/ group. B The fluorescence intensity of G. parasuis-exosome (20 mg/kg) or Mock-exosome (20 mg/kg) in different organs of mice. n = 6 mice/group. C–E The effect of exosomes and circHIF1α on the survival of mice after G. parasuis (C)/SS2 (D)/S. aureus (E) infection. n = 6 mice/group. F–H The effect of exo somes and circHIF1α on G. parasuis (F)/SS2 (G)/S. aureus (H) count of different organs after mice were infected with G. parasuis. I H&E staining of various organs after mice were infected with G. parasuis followed treating with exosome or circHIF1α. Scale bar, 50 μm. n = 6 mice/group. Data are represented as mean ± SD. *p < 0.05, **p < 0.01

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Infection, In Vivo, Fluorescence, Staining

Journal: Journal of nanobiotechnology

Article Title: m6A-modified exosome-derived circHIF1α binding to KH domain of IGF2BP3 mediates DNA damage and arrests G1/S transition phase to resists bacterial infection in bacteremia.

doi: 10.1186/s12951-024-02932-4

Figure Lengend Snippet: Fig. 9 Proposed model for the potential function of exosomal circHIF1α in bacterial infection progression

Article Snippet: In the immunofluorescence assay, PK-15 cells were transiently transfected with Cy3labelled circHIF1α and proceeded to FISH assay, and then incubated with anti-IGF2BP3 antibody (Proteintech, Chicago, USA) to observe the colocalization of circHIF1α and IGF2BP3.

Techniques: Infection

Journal: Cell biology and toxicology

Article Title: Downregulation of MYBL1 in endothelial cells contributes to atherosclerosis by repressing PLEKHM1-inducing autophagy.

doi: 10.1007/s10565-024-09873-6

Figure Lengend Snippet: Fig. 9 A schematic diagram for the underlying mechanism of the UPRmt in regulating IVDD. OxLDL attacked MYBL1 to induce apoptosis of endothelial cells in the atheroscle- rosis. PLEKHM1, as a downstream of MYBL1 in endothelial cells, promoted the fusion of autophagy and lysosomes in endothelial cells

Article Snippet: The primary antibodies were listed as follow: MYBL1 (Affinity, AF9007, 1:1000), PLEKHM1 (Proteintech, 16202–1-AP, 1:1000), β-actin (Proteintech, 81115–1-RR, 1:5000), AMPK (Proteintech, 10929–2- AP, 1: 1000), GABARAP (Proteintech, 18723–1-AP, 1:500), Cleaved-Caspase3 (CST, 9664, 1:1000), Bcl-2 (Proteintech, 26593–1-AP, 1:1000), Bax (Proteintech, 50599–2-Ig, 1:1000), hVps41 (Proteintech, 13869–1-AP, 1:500), hVps11 (Proteintech, 19140–1-AP, 1:500), Rab7 (Proteintech, 55469–1-AP, 1:500), LC3 (Novus, NB1002220, 1:1000) and p62 (CST, 88588, 1:1000).

Techniques:

Journal: PLoS ONE

Article Title: Novel Nonphosphorylated Peptides with Conserved Sequences Selectively Bind to Grb7 SH2 Domain with Affinity Comparable to Its Phosphorylated Ligand

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: Nonphosphorylated peptides interacting with the Grb7 SH2 domain (abridged sequences of conserved amino acid sequences).

Article Snippet: The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques:

Journal: PLoS ONE

Article Title: Novel Nonphosphorylated Peptides with Conserved Sequences Selectively Bind to Grb7 SH2 Domain with Affinity Comparable to Its Phosphorylated Ligand

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: A ) Expression of nonphosphorylated peptides. B ) Interactions between nonphosphorylated peptides and the Grb7 SH2 domain.

Article Snippet: The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Novel Nonphosphorylated Peptides with Conserved Sequences Selectively Bind to Grb7 SH2 Domain with Affinity Comparable to Its Phosphorylated Ligand

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: A ) Different concentrations of synthesized phosphorylated ligands pY (1180) (DEEYEpY(1180)MNRRR) from ErbB3 and the nonphosphorylated ligands 41-A (VAVGIPTQPTTSSEPSPPSNPPWDPGRV) from the nonphosphorylated ligand 41 were added respectively into the interacting complexes of Grb7 SH2 domain and peptide 41 (on the top). The bands were scanned and IC 50 values were calculated (on the bottom). B ) Structural modeling for the nonphosphorylated peptide 41-A (VAV GIPTQPTTSSEPSPPSNPPWDPGRV) binding to the Grb7 SH2 domain was performed with MOE. Left was the structural model of the Grb7 SH2/pY (1139) of ErbB2 , right was that of the Grb7 SH2/41-A. Red section represented α helix; yellow represented β strand; white represented loop in the Grb7 SH2 domain; green represented the binding ligand; for the nonphosphorylated peptide, left was the N-terminus, and right was the C-terminus.

Article Snippet: The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques: Synthesized, Binding Assay

Journal: PLoS ONE

Article Title: Novel Nonphosphorylated Peptides with Conserved Sequences Selectively Bind to Grb7 SH2 Domain with Affinity Comparable to Its Phosphorylated Ligand

doi: 10.1371/journal.pone.0029902

Figure Lengend Snippet: A ) Protein expression. The endogenous Grb7 protein expression (left) and expression of the fusion protein of 41-mRLUC-3×Flag (right) in the SK-BR-3 breast cancer cells were identified by western blot. B ) SK-BR-3 breast cancer cell proliferation analysis. Double-factor variance analysis was performed to analyze the differences between the adjusted absorbance and different transfected cell groups, and between the adjusted absorbance and different time points. Statistically significant differences existed in both of groups (n = 10, p<0.001).

Article Snippet: The ABL_1 and Grb2 plasmids were gifts from Professor Ruibao Ren (Brandeis University, USA), and the Grb7 (BC006535) and Grb14 cDNA (BC053559) plasmids were from the Proteintech Group.

Techniques: Expressing, Western Blot, Transfection