Journal: Journal of Cardiothoracic Surgery

Article Title: NSUN2 inhibits NCOA4 expression to alleviate ferroptosis and inflammation in sepsis-induced myocardial injury in a m 5 C manner

doi: 10.1186/s13019-025-03554-z

Figure Lengend Snippet: NSUN2 suppressed ferroptosis in LPS-induced SIMI. Cellular A , Fe 2 + level, B , GSH content, and C , relative ROS production in each group were determined by commercial kits; D , Western blot was performed to assess the protein levels of NCOA4, FTH1, and GPX4 in each group Fe 2+ , ferrous iron; GSH, glutathione; ROS, reactive oxygen species; NCOA, nuclear receptor coactivator; FTH1, ferritin heavy chain; GPX, glutathione peroxidase

Article Snippet: The primary antibodies used were rabbit antibodies specific for NCOA4 (1:1000; ab314553; Abcam, Cambridge, MA, USA), FTH1 (1:1000; CSB- PA008485 ; Cusabio Engineering Co., LTD, Wuhan, China), GPX4 (1:1000; ab125066; Abcam), and GAPDH (1:1000; ab8245; Abcam).

Techniques: Western Blot

Journal: Journal of Cardiothoracic Surgery

Article Title: NSUN2 inhibits NCOA4 expression to alleviate ferroptosis and inflammation in sepsis-induced myocardial injury in a m 5 C manner

doi: 10.1186/s13019-025-03554-z

Figure Lengend Snippet: NSUN2 inhibited NCOA4 expression in a m 5 C-dependent manner. A , The expression of NCOA4, FTH1, and GPX4 after NSUN2 overexpression in H9c2 cells was detected by RT-qPCR; B , MeRIP-qPCR assay was performed to detect m 5 C levels of NCOA4, FTH1, and GPX4 in vector and NSUN2 groups; C , RIP assay evaluated the interaction of NSUN2 and NCOA4 in H9c2 cells; D , Random Forest method was used to predict possible m 5 C sites of NCOA4; Dual-luciferase gene reporter assay evaluated the binding of NSUN2 and NCOA4 at sites E , 501, F , 1138, and G , 2917; H , RNA stability assay was used to detect the NCOA4 expression when actinomycin D treated at different time points (0, 4, 8, and 12 h) after NSUN2 overexpression NCOA, nuclear receptor coactivator; FTH1, ferritin heavy chain; GPX, glutathione peroxidase; NSUN, NOL1/NOP2/SUN domain; RT-qPCR, reverse transcription-polymerase chain reaction; MeRIP, Methylated RNA immunoprecipitation; RIP, RNA immunoprecipitation; m 5 C, 5-methylcytosine

Article Snippet: The primary antibodies used were rabbit antibodies specific for NCOA4 (1:1000; ab314553; Abcam, Cambridge, MA, USA), FTH1 (1:1000; CSB- PA008485 ; Cusabio Engineering Co., LTD, Wuhan, China), GPX4 (1:1000; ab125066; Abcam), and GAPDH (1:1000; ab8245; Abcam).

Techniques: Expressing, Over Expression, Quantitative RT-PCR, Plasmid Preparation, Luciferase, Reporter Assay, Binding Assay, Stability Assay, Reverse Transcription, Polymerase Chain Reaction, Methylation, RNA Immunoprecipitation

Journal: Journal of Cardiothoracic Surgery

Article Title: NSUN2 inhibits NCOA4 expression to alleviate ferroptosis and inflammation in sepsis-induced myocardial injury in a m 5 C manner

doi: 10.1186/s13019-025-03554-z

Figure Lengend Snippet: Overexpressing NCOA4 downregulated cell viability and upregulated LDH activity, inflammation, and ferroptosis in LPS-induced SIMI. A , NCOA4 mRNA level was detected by RT-qPCR; B , The cell viability of H9c2 cells in each group was analyzed by CCK-8 assay; C , Determination of LDH enzyme activity in each group; D , The contents of TNF-α, IL-6, and IL-8 in each group was evaluated by ELISA; Cellular E , Fe 2 + level, F , GSH content, and G , relative ROS production in each group were determined by commercial kits; H , The protein levels of NCOA4, FTH1, and GPX4 in each group was analyzed by Western blot RT-qPCR, reverse transcription-polymerase chain reaction; CCK-8, cell counting kit-8; LDH, lactate dehydrogenase; ELISA, enzyme-linked immunosorbent assay; TNF-α, tumor necrosis factor-α; IL, interleukin; Fe 2+ , ferrous iron; GSH, glutathione; ROS, reactive oxygen species; NCOA, nuclear receptor coactivator; FTH1, ferritin heavy chain; GPX, glutathione peroxidase

Article Snippet: The primary antibodies used were rabbit antibodies specific for NCOA4 (1:1000; ab314553; Abcam, Cambridge, MA, USA), FTH1 (1:1000; CSB- PA008485 ; Cusabio Engineering Co., LTD, Wuhan, China), GPX4 (1:1000; ab125066; Abcam), and GAPDH (1:1000; ab8245; Abcam).

Techniques: Activity Assay, Quantitative RT-PCR, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Cell Counting

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Evolution of infection by Mycobacterium avium in mice deficient in H-ferritin in myeloid cells. Fth1 +/+ (blue) and Fth1 −/− (red) mice were intravenously infected with 10 6 CFU of M. avium 25291. ( A ) Animals’ body weight throughout the experiment. Data are expressed as percentage of the initial weight, presented as the mean ± SD of 5 to 8 animals per group. Empty circles: noninfected; filled circles: infected. ( B ) Bacterial burden 60 days post-infection in the liver, spleen, and bone marrow of 8 Fth1 +/+ and 7 Fth1 −/− mice. Bars represent the mean, and circles represent each animal. ( C ) Kaplan–Meier survival curve corresponding to 12 Fth1 +/+ and 16 Fth1 −/− animals. Mice were euthanized when they lost 20% of the initial weight (between day 43 and day 152 post infection). ( D ) Bacterial burden in the liver, spleen, and bone marrow, at the time of euthanasia in the survival curve, of 12 Fth1 +/+ and 16 Fth1 −/− mice. Bars represent the mean and circles represent each animal. Statistics: multiple t -test in ( A , B , D ); log-rank (Mantel–Cox) test in ( C ). ** p < 0.01 and *** p < 0.001 when comparing Fth1 −/− with Fth1 +/+ mice.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Inflammatory response in mice infected with M. avium . Fth1 +/+ (blue) and Fth1 −/− (red) mice were intravenously infected with 10 6 CFU of M. avium 25291 and euthanized 60 days later. ( A ) TNF-alpha, ( B ) IFN-gamma, and ( C ) IL-6 were measured in the serum, using a cytometric bead array kit. Bars represent the mean, and the circles represent individual values of 4 to 8 animals per group. Statistics: two-way ANOVA followed by Sidak multiple-comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 when comparing Fth1 +/+ to Fth1 −/− mice. ## p < 0.01, ### p < 0.001 when comparing infected to noninfected mice of the same genotype. ( D – G ) Representative images of H&E-stained liver sections from noninfected (NI) and infected (60 dpi) Fth1 +/+ and Fth1 −/− mice. Representative granulomas are outlined. Scale bar: 200 µm.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection, Comparison, Staining

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Impact of infection on iron distribution. Sera from Fth1 +/+ (blue) and Fth1 −/− (red) mice infected for 60 days with M. avium 25291 (60 dpi) or from noninfected mice (NI), were used to measure ( A ) iron, ( B ) transferrin saturation, and ( C ) total ferritin ( n = 5 to 8). ( D , E ) Serum FTL and FTH1 subunits were measured by ELISA ( n = 2 to 4). ( F ) Liver sections were collected, and the iron concentration was determined by atomic absorption spectrometry ( n = 5 to 8). Bars represent the mean, and circles represent each animal. Statistics: two-way ANOVA followed by Sidak multiple-comparison post hoc test. ** p < 0.01, *** p < 0.001 when comparing Fth1 −/− with Fth1 +/+ mice in the same experimental condition. # p < 0.05, ## p < 0.01, ### p < 0.001 when comparing infected to noninfected mice of the same genotype. ( G – J ) Perls’ staining of liver sections was used to evaluate iron distribution. Arrows indicate regions of iron accumulation (blue staining). Scale bar = 200 µm.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Concentration Assay, Comparison, Staining

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Effect of iron overload on the growth of M. avium in the liver. Groups of Fth1 +/+ (blue) and Fth1 −/− (red) mice were intraperitoneally injected with 10 mg of iron, in the form of iron–dextran, 10 days before intravenous infection with 10 6 CFU of M. avium 25291. Sixty days after infection, noninfected (NI) and infected (60 dpi) mice were euthanized. ( A ) Liver iron was quantified by atomic absorption spectrometry ( n = 4 to 8 per group). ( B ) Bacterial burden in the livers, 60 days after infection ( n = 6 to 8 per group). The bars represent the mean of the group, and each circle corresponds to one animal. Statistics: two-way ANOVA followed by Sidak multiple-comparison post hoc test. *** p < 0.001 when comparing Fth1 −/− to Fth1 +/+ mice in the same experimental condition. @ p < 0.001 when comparing iron-overloaded with non-iron-overloaded mice of the same genotype and infection status.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Injection, Infection, Comparison

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Iron-related gene expression induced by mycobacterial infection.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Gene Expression, Infection

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Effects of infection and myeloid Fth1 deficiency in iron and ferroportin distribution in the liver. Liver sections were obtained from ( A , B ) noninfected mice or ( C – H ) mice infected with M. avium for 60 days. Sections were stained for ( A – D , G , H ) ferroportin, using immunohistochemistry (brown staining) or for ( E , F ) iron, using the Perls’ method (blue staining). ( E – H ) Mice were intraperitoneally injected with 10 mg of iron, as iron–dextran, 10 days before infection. Representative images of at least three animals per experimental condition. Representative granulomas are outlined. Scale bar: 200 µm.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection, Staining, Immunohistochemistry, Injection

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Primers sequences.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Sequencing