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GAL‐9 protein is an important regulatory molecule in neutrophil hyperactivity. (A) Intracellular GAL‐9 protein expression of circulating neutrophils in the sham group and 80 days post‐radiation. n = 6. (B) Circulating GAL‐9 protein level by ELISA kits. n = 8. (C, D) (C) Scheme and (D) the IFN‐γ expression of circulating neutrophils in the sham group under different conditions. n = 3. (E–M) (E) Scheme of the IFN‐γ and NETs expression of circulating neutrophils in the sham group by treatment with (F, H, K) circulating serum at 80 days post‐radiation and (F, I, L) culture supernatant of GAL‐9 high neutrophils. (F, J, M) IFN‐γ and NETs expression of GAL‐9 high neutrophils 80 days post‐radiation under different stimuli. n = 4–5. (N–Q) Assessment of the effect of GAL‐9 protein on the polarization of bone marrow macrophages under different conditions. (N) The scheme and the effect of (O) rmGAL‐9 protein, (P) culture supernatant of GAL‐9 high neutrophils, and (Q) circulating serum at 80 days post‐radiation were shown. n = 3. (R–S) Representative plots and statistics of bone marrow <t>CD47</t> + neutrophils in the sham group and 80 days post‐radiation. n = 4. (T‐BB) Assessment of the reversal effect of GAL‐9 intervention in mice. (T) Scheme of the administration of anti‐GAL‐9 and rmGAL‐9 proteins in the local radiation group and the sham group, respectively. (U) Circulating GAL‐9 high neutrophils, bone marrow (V) CMP cells, (W) CLP cells, (X) non‐immune cells, (Y) macrophages, and (Z, AA) their polarization state and (BB) frailty index score were shown after different treatments in the local radiation group and the sham group. n = 3–5. Data are presented as mean ± SD; each dot represents an individual animal from at least 2–4 independent experiments that used male and female mice. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical analyses were performed using unpaired Student's t ‐test (A, B, S, U), one‐way ANOVA (D, G–M, V–AA), and two‐way ANOVA (O–Q).
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GAL‐9 protein is an important regulatory molecule in neutrophil hyperactivity. (A) Intracellular GAL‐9 protein expression of circulating neutrophils in the sham group and 80 days post‐radiation. n = 6. (B) Circulating GAL‐9 protein level by ELISA kits. n = 8. (C, D) (C) Scheme and (D) the IFN‐γ expression of circulating neutrophils in the sham group under different conditions. n = 3. (E–M) (E) Scheme of the IFN‐γ and NETs expression of circulating neutrophils in the sham group by treatment with (F, H, K) circulating serum at 80 days post‐radiation and (F, I, L) culture supernatant of GAL‐9 high neutrophils. (F, J, M) IFN‐γ and NETs expression of GAL‐9 high neutrophils 80 days post‐radiation under different stimuli. n = 4–5. (N–Q) Assessment of the effect of GAL‐9 protein on the polarization of bone marrow macrophages under different conditions. (N) The scheme and the effect of (O) rmGAL‐9 protein, (P) culture supernatant of GAL‐9 high neutrophils, and (Q) circulating serum at 80 days post‐radiation were shown. n = 3. (R–S) Representative plots and statistics of bone marrow <t>CD47</t> + neutrophils in the sham group and 80 days post‐radiation. n = 4. (T‐BB) Assessment of the reversal effect of GAL‐9 intervention in mice. (T) Scheme of the administration of anti‐GAL‐9 and rmGAL‐9 proteins in the local radiation group and the sham group, respectively. (U) Circulating GAL‐9 high neutrophils, bone marrow (V) CMP cells, (W) CLP cells, (X) non‐immune cells, (Y) macrophages, and (Z, AA) their polarization state and (BB) frailty index score were shown after different treatments in the local radiation group and the sham group. n = 3–5. Data are presented as mean ± SD; each dot represents an individual animal from at least 2–4 independent experiments that used male and female mice. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical analyses were performed using unpaired Student's t ‐test (A, B, S, U), one‐way ANOVA (D, G–M, V–AA), and two‐way ANOVA (O–Q).
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GAL‐9 protein is an important regulatory molecule in neutrophil hyperactivity. (A) Intracellular GAL‐9 protein expression of circulating neutrophils in the sham group and 80 days post‐radiation. n = 6. (B) Circulating GAL‐9 protein level by ELISA kits. n = 8. (C, D) (C) Scheme and (D) the IFN‐γ expression of circulating neutrophils in the sham group under different conditions. n = 3. (E–M) (E) Scheme of the IFN‐γ and NETs expression of circulating neutrophils in the sham group by treatment with (F, H, K) circulating serum at 80 days post‐radiation and (F, I, L) culture supernatant of GAL‐9 high neutrophils. (F, J, M) IFN‐γ and NETs expression of GAL‐9 high neutrophils 80 days post‐radiation under different stimuli. n = 4–5. (N–Q) Assessment of the effect of GAL‐9 protein on the polarization of bone marrow macrophages under different conditions. (N) The scheme and the effect of (O) rmGAL‐9 protein, (P) culture supernatant of GAL‐9 high neutrophils, and (Q) circulating serum at 80 days post‐radiation were shown. n = 3. (R–S) Representative plots and statistics of bone marrow <t>CD47</t> + neutrophils in the sham group and 80 days post‐radiation. n = 4. (T‐BB) Assessment of the reversal effect of GAL‐9 intervention in mice. (T) Scheme of the administration of anti‐GAL‐9 and rmGAL‐9 proteins in the local radiation group and the sham group, respectively. (U) Circulating GAL‐9 high neutrophils, bone marrow (V) CMP cells, (W) CLP cells, (X) non‐immune cells, (Y) macrophages, and (Z, AA) their polarization state and (BB) frailty index score were shown after different treatments in the local radiation group and the sham group. n = 3–5. Data are presented as mean ± SD; each dot represents an individual animal from at least 2–4 independent experiments that used male and female mice. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical analyses were performed using unpaired Student's t ‐test (A, B, S, U), one‐way ANOVA (D, G–M, V–AA), and two‐way ANOVA (O–Q).
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GAL‐9 protein is an important regulatory molecule in neutrophil hyperactivity. (A) Intracellular GAL‐9 protein expression of circulating neutrophils in the sham group and 80 days post‐radiation. n = 6. (B) Circulating GAL‐9 protein level by ELISA kits. n = 8. (C, D) (C) Scheme and (D) the IFN‐γ expression of circulating neutrophils in the sham group under different conditions. n = 3. (E–M) (E) Scheme of the IFN‐γ and NETs expression of circulating neutrophils in the sham group by treatment with (F, H, K) circulating serum at 80 days post‐radiation and (F, I, L) culture supernatant of GAL‐9 high neutrophils. (F, J, M) IFN‐γ and NETs expression of GAL‐9 high neutrophils 80 days post‐radiation under different stimuli. n = 4–5. (N–Q) Assessment of the effect of GAL‐9 protein on the polarization of bone marrow macrophages under different conditions. (N) The scheme and the effect of (O) rmGAL‐9 protein, (P) culture supernatant of GAL‐9 high neutrophils, and (Q) circulating serum at 80 days post‐radiation were shown. n = 3. (R–S) Representative plots and statistics of bone marrow <t>CD47</t> + neutrophils in the sham group and 80 days post‐radiation. n = 4. (T‐BB) Assessment of the reversal effect of GAL‐9 intervention in mice. (T) Scheme of the administration of anti‐GAL‐9 and rmGAL‐9 proteins in the local radiation group and the sham group, respectively. (U) Circulating GAL‐9 high neutrophils, bone marrow (V) CMP cells, (W) CLP cells, (X) non‐immune cells, (Y) macrophages, and (Z, AA) their polarization state and (BB) frailty index score were shown after different treatments in the local radiation group and the sham group. n = 3–5. Data are presented as mean ± SD; each dot represents an individual animal from at least 2–4 independent experiments that used male and female mice. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical analyses were performed using unpaired Student's t ‐test (A, B, S, U), one‐way ANOVA (D, G–M, V–AA), and two‐way ANOVA (O–Q).
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GAL‐9 protein is an important regulatory molecule in neutrophil hyperactivity. (A) Intracellular GAL‐9 protein expression of circulating neutrophils in the sham group and 80 days post‐radiation. n = 6. (B) Circulating GAL‐9 protein level by ELISA kits. n = 8. (C, D) (C) Scheme and (D) the IFN‐γ expression of circulating neutrophils in the sham group under different conditions. n = 3. (E–M) (E) Scheme of the IFN‐γ and NETs expression of circulating neutrophils in the sham group by treatment with (F, H, K) circulating serum at 80 days post‐radiation and (F, I, L) culture supernatant of GAL‐9 high neutrophils. (F, J, M) IFN‐γ and NETs expression of GAL‐9 high neutrophils 80 days post‐radiation under different stimuli. n = 4–5. (N–Q) Assessment of the effect of GAL‐9 protein on the polarization of bone marrow macrophages under different conditions. (N) The scheme and the effect of (O) rmGAL‐9 protein, (P) culture supernatant of GAL‐9 high neutrophils, and (Q) circulating serum at 80 days post‐radiation were shown. n = 3. (R–S) Representative plots and statistics of bone marrow CD47 + neutrophils in the sham group and 80 days post‐radiation. n = 4. (T‐BB) Assessment of the reversal effect of GAL‐9 intervention in mice. (T) Scheme of the administration of anti‐GAL‐9 and rmGAL‐9 proteins in the local radiation group and the sham group, respectively. (U) Circulating GAL‐9 high neutrophils, bone marrow (V) CMP cells, (W) CLP cells, (X) non‐immune cells, (Y) macrophages, and (Z, AA) their polarization state and (BB) frailty index score were shown after different treatments in the local radiation group and the sham group. n = 3–5. Data are presented as mean ± SD; each dot represents an individual animal from at least 2–4 independent experiments that used male and female mice. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical analyses were performed using unpaired Student's t ‐test (A, B, S, U), one‐way ANOVA (D, G–M, V–AA), and two‐way ANOVA (O–Q).

Journal: Aging Cell

Article Title: Galectin‐9 high Neutrophils Exacerbate Radiation‐Induced Frailty

doi: 10.1111/acel.70448

Figure Lengend Snippet: GAL‐9 protein is an important regulatory molecule in neutrophil hyperactivity. (A) Intracellular GAL‐9 protein expression of circulating neutrophils in the sham group and 80 days post‐radiation. n = 6. (B) Circulating GAL‐9 protein level by ELISA kits. n = 8. (C, D) (C) Scheme and (D) the IFN‐γ expression of circulating neutrophils in the sham group under different conditions. n = 3. (E–M) (E) Scheme of the IFN‐γ and NETs expression of circulating neutrophils in the sham group by treatment with (F, H, K) circulating serum at 80 days post‐radiation and (F, I, L) culture supernatant of GAL‐9 high neutrophils. (F, J, M) IFN‐γ and NETs expression of GAL‐9 high neutrophils 80 days post‐radiation under different stimuli. n = 4–5. (N–Q) Assessment of the effect of GAL‐9 protein on the polarization of bone marrow macrophages under different conditions. (N) The scheme and the effect of (O) rmGAL‐9 protein, (P) culture supernatant of GAL‐9 high neutrophils, and (Q) circulating serum at 80 days post‐radiation were shown. n = 3. (R–S) Representative plots and statistics of bone marrow CD47 + neutrophils in the sham group and 80 days post‐radiation. n = 4. (T‐BB) Assessment of the reversal effect of GAL‐9 intervention in mice. (T) Scheme of the administration of anti‐GAL‐9 and rmGAL‐9 proteins in the local radiation group and the sham group, respectively. (U) Circulating GAL‐9 high neutrophils, bone marrow (V) CMP cells, (W) CLP cells, (X) non‐immune cells, (Y) macrophages, and (Z, AA) their polarization state and (BB) frailty index score were shown after different treatments in the local radiation group and the sham group. n = 3–5. Data are presented as mean ± SD; each dot represents an individual animal from at least 2–4 independent experiments that used male and female mice. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001. Statistical analyses were performed using unpaired Student's t ‐test (A, B, S, U), one‐way ANOVA (D, G–M, V–AA), and two‐way ANOVA (O–Q).

Article Snippet: Primary antibodies used were Gal‐9 (1:500, Abcam #ab69630), Myeloperoxidase (1:50, Abcam #ab90810), Histone H3 (1:1000, Abcam #ab281584), Ly6g + Ly6c (1:500, Abcam #ab25377), and CD47 (1:500, Abmart #T55251S).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay