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