Journal: Journal of Dental Sciences

Article Title: Fatostatin delayed lip sensory recovery after inferior alveolar nerve transection by inhibiting sterol regulatory element-binding protein 1

doi: 10.1016/j.jds.2025.04.024

Figure Lengend Snippet: Fatostatin impaired lip sensory recovery following inferior alveolar nerve transection in vivo . (A) Quantitative sensory testing using Von Frey filaments showed the sensory recovery of the lower lip in the control and fatostatin-treated groups, measured every 4 days for 40 days. The results show that fatostatin significantly delayed lip sensory recovery compared to the control group. (B) Quantitative real-time PCR analysis of gene expression in the trigeminal ganglia demonstrated that fatostatin treatment significantly decreased the mRNA levels of SREBP1, FASN, ACLY, ATF3, and NGF. (C) Western blot analysis confirmed that fatostatin treatment reduced the expression levels of SREBP1, ACLY, and ATF3 in the trigeminal ganglia compared to the control group. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. SREBP1, sterol regulatory element-binding protein 1; FASN, fatty acid synthase; ACLY, ATP citrate lyase; ATF3, activating transcription factor 3; NGF, nerve growth factor.

Article Snippet: The used primary antibodies included SREBP1 (sc-13551, Santa Cruz, Dallas, TX, USA), ACLY (sc-517267, Santa Cruz), ATF3 (18665, Cell Signaling Technology, Boston, MA, USA), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vivo, Control, Real-time Polymerase Chain Reaction, Gene Expression, Western Blot, Expressing, Binding Assay

Journal: Journal of Dental Sciences

Article Title: Fatostatin delayed lip sensory recovery after inferior alveolar nerve transection by inhibiting sterol regulatory element-binding protein 1

doi: 10.1016/j.jds.2025.04.024

Figure Lengend Snippet: Fatostatin inhibits axonal growth of primary trigeminal neurons in vitro . (A) Images of cultured primary neurons stained with NF200. The upper panels show the control group (left) and the fatostatin-treated group (right) at 20 × magnification. The lower panels show higher magnification (100 × ) of neurons in both groups. Scale bars in the upper panels indicate 100 μm, and in the lower panels, 20 μm. (B) Quantitative analysis of axonal length in the top 50 neurons, showing that fatostatin significantly reduced axonal growth compared to the control group. (C) Quantitative PCR analysis of key genes related to lipid metabolism and axonal regeneration, including SREBP1, FASN, ACLY, ATF3, and NGF. The results show that fatostatin treatment significantly reduced the expression levels of these genes, with SREBP1 and FASN showing the most prominent changes. (D) Western blot analysis of SREBP1, ACLY, and ATF3 protein expression levels, demonstrating that fatostatin treatment decreased SREBP1, ACLY, and ATF3 protein levels compared to control. GAPDH was used as the loading control. ∗∗∗, P < 0.001, ∗∗∗∗, P < 0.0001. SREBP1, sterol regulatory element-binding protein 1; FASN, fatty acid synthase; ACLY, ATP citrate lyase; ATF3, activating transcription factor 3; NGF, nerve growth factor.

Article Snippet: The used primary antibodies included SREBP1 (sc-13551, Santa Cruz, Dallas, TX, USA), ACLY (sc-517267, Santa Cruz), ATF3 (18665, Cell Signaling Technology, Boston, MA, USA), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vitro, Cell Culture, Staining, Control, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Binding Assay

Journal: Journal of Dental Sciences

Article Title: Serotonin promotes lip sensory recovery after inferior alveolar nerve transection via histone serotonylation

doi: 10.1016/j.jds.2025.04.001

Figure Lengend Snippet: Levels of serotonin production and histone serotonylation were improved in trigeminal ganglia after inferior alveolar nerve transection. (A) Photos of inferior alveolar nerve transection mice model. The triangle in (a1 shows the exposure of the inferior alveolar nerve. The asterisk in (a2) shows the transection of the inferior alveolar nerve. (B) Quantitative sensory testing with Von Frey filaments showed the lower lip sensation after sham surgery (Control group) versus inferior alveolar nerve transection (Transected group) in mice, measured every four days for 8 weeks (n = 5). (C) ELISA showed the serotonin level in trigeminal ganglia (n = 5). (D) Quantitative real-time polymerase chain reaction showed the genes related to serotonin production (including TGM2, MAOA, MAOB, HTR1A, HTR2A, and HTR3A) in trigeminal ganglia (n = 5). (E) Immunofluorescence showed the level of H3Q5Ser, H3K4me3, Tuj in the trigeminal ganglia. Bar indicated 20 μm. (F) Quantitative analysis of immunofluorescence strength of H3Q5Ser and H3K4me3 in Tuj-positive area (n = 5). (G) Western blot showed the level of H3Q5Ser and H3K4me3 in the trigeminal ganglia. (H) Quantitative analysis of the relative ratio of H3Q5Ser per H3K4me3 in the blots (n = 3). (I) Western blot showed the protein expression level of TGM2 in the trigeminal ganglia. (J) Quantitative analysis of the relative ratio of TGM2 per GAPDH in the blots (n = 3). ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001. TGM2, transglutaminase 2; MAOA, monoamine oxidase A; MAOB, monoamine oxidase B; HTR1A, 5-hydroxytryptamine receptor 1A; HTR2A, 5-hydroxytryptamine receptor 2A; HTR3A, 5-hydroxytryptamine receptor 3A; H3Q5Ser, histone H3 glutamine 5 serotonylation; H3K4me3, histone H3 lysine 4 trimethylation; Tuj1, neuron-specific class III beta-tubulin; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The used primary antibodies included H3Q5ser (PTM-1420, PTM Bio, Hangzhou, China), H3K4me3 (PTM-5019, PTM Bio), H3 (PTM-6613, PTM Bio), TGM2 (3557, Cell Signaling Technology, Boston, MA, USA), ATF3 (18665, Cell Signaling Technology), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: Control, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Immunofluorescence, Western Blot, Expressing

Journal: Journal of Dental Sciences

Article Title: Serotonin promotes lip sensory recovery after inferior alveolar nerve transection via histone serotonylation

doi: 10.1016/j.jds.2025.04.001

Figure Lengend Snippet: Exogenuous serotonin improved the axonal length of primary neurons of trigeminal ganglia via histone serotonylation in vitro . (A) Images of Cultured primary neurons. (a1) and (a2) were cells in the control group. (b1) and (b2) were cells in the serotonin group (addition of 100 μM Serotonin hydrochloride). Bar in (a1) and (b1) indicated 100 μm. Bar in (a2) and (b2) indicated 20 μm. (B) Quantitative analysis of axon length of top 50 neurons in the images. (C) Western blot showed the level of H3Q5Ser and H3K4me3 in the neurons. (D) Quantitative analysis of the relative ratio of H3Q5Ser per H3K4me3 in the blots (n = 3). (E) Quantitative real-time polymerase chain reaction showed the genes related to axon regeneration (including NGF, ATF3, NTF3, NTF5, and GDNF) and lipid metabolism (FASN, SREBF1, and APOD) in the neurons (n = 5). (F) Western blot showed the protein expression level of ATF3 in the trigeminal ganglia. (G) Quantitative analysis of the relative ratio of ATF3 per GAPDH in the blots (n = 3). (H) ELISA showed the NGF level in the nerons (n = 5). ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001. NGF, nerve growth factor; ATF3, activating transcription factor 3; NTF3, neurotrophin-3; NTF5, neurotrophin-5; GDNF, glial cell line-derived neurotrophic factor; FASN, fatty acid synthase; SREBF1, sterol regulatory element-binding transcription factor 1; APOD, apolipoprotein D; H3Q5Ser, histone H3 glutamine 5 serotonylation; H3K4me3, histone H3 lysine 4 trimethylation; GAPDH, glyceraldehyde-3-phosphate dehydrogenase.

Article Snippet: The used primary antibodies included H3Q5ser (PTM-1420, PTM Bio, Hangzhou, China), H3K4me3 (PTM-5019, PTM Bio), H3 (PTM-6613, PTM Bio), TGM2 (3557, Cell Signaling Technology, Boston, MA, USA), ATF3 (18665, Cell Signaling Technology), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vitro, Cell Culture, Control, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Binding Assay

Journal: Journal of Dental Sciences

Article Title: Serotonin promotes lip sensory recovery after inferior alveolar nerve transection via histone serotonylation

doi: 10.1016/j.jds.2025.04.001

Figure Lengend Snippet: Exogenous serotonin improved the lip sensory recovery after inferior alveolar nerve transection via histone serotonylation in vivo . (A) Quantitative sensory testing with Von Frey filaments showed the lower lip sensation after transection alone (Trans group) versus transection plus exogenous serotonin group (administration of 100 μM Serotonin hydrochloride) (Trans + Ser group) in mice, measured every four days for 8 weeks (n = 5). (B) Immunofluorescence showed the level of H3Q5Ser, H3K4me3, Tuj in the trigeminal ganglia. Bar indicated 20 μm. (C) Quantitative analysis of immunofluorescence strength of H3Q5Ser and H3K4me3 in Tuj-positive area (n = 5). (D) Western blot showed the level of H3Q5Ser and H3K4me3 in the trigeminal ganglia. (E) Quantitative analysis of the relative ratio of H3Q5Ser per H3K4me3 in the blots (n = 3). (F) Quantitative real-time polymerase chain reaction showed the genes related to axon regeneration (including NGF, ATF3, NTF3, NTF5, and GDNF) and lipid metabolism (FASN, SREBF1, and APOD) in the trigeminal ganglia (n = 5). (G) Western blot showed the protein expression level of ATF3 in the trigeminal ganglia. (H) Quantitative analysis of the relative ratio of ATF3 per GAPDH in the blots (n = 3). (I) ELISA showed the NGF level in the trigeminal ganglia (n = 5). ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001. NGF, nerve growth factor; ATF3, activating transcription factor 3; NTF3, neurotrophin-3; NTF5, neurotrophin-5; GDNF, glial cell line-derived neurotrophic factor; FASN, fatty acid synthase; SREBF1, sterol regulatory element-binding transcription factor 1; APOD, apolipoprotein D; H3Q5Ser, histone H3 glutamine 5 serotonylation; H3K4me3, histone H3 lysine 4 trimethylation; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Tuj1, neuron-specific class III β-tubulin.

Article Snippet: The used primary antibodies included H3Q5ser (PTM-1420, PTM Bio, Hangzhou, China), H3K4me3 (PTM-5019, PTM Bio), H3 (PTM-6613, PTM Bio), TGM2 (3557, Cell Signaling Technology, Boston, MA, USA), ATF3 (18665, Cell Signaling Technology), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vivo, Immunofluorescence, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Binding Assay

Journal: Journal of Dental Sciences

Article Title: Serotonin promotes lip sensory recovery after inferior alveolar nerve transection via histone serotonylation

doi: 10.1016/j.jds.2025.04.001

Figure Lengend Snippet: GK921 inhibited the serotonin-induced axonal growth of primary neurons of trigeminal ganglia by reducing histone serotonylation in vitro . (A) Images of Cultured primary neurons. (a1) and (a2) were cells in the serotonin (Ser) group (addition of 100 μM Serotonin hydrochloride). (b1) and (b2) were cells in the serotonin + GK921 (Ser + GK921) group (addition of 100 μM Serotonin hydrochloride plus 50 μM GK921). Bar in (a1) and (b1) indicated 100 μm. Bar in (a2) and (b2) indicated 20 μm. (B) Quantitative analysis of axon length of top 50 neurons in the images. (C) Western blot showed the level of H3Q5Ser and H3K4me3 in the neurons. (D) Quantitative analysis of the relative ratio of H3Q5Ser per H3K4me3 in the blots (n = 3). (E) Quantitative real-time polymerase chain reaction showed the genes related to axon regeneration (including NGF, ATF3, NTF3, NTF5, and GDNF) and lipid metabolism (FASN, SREBF1, and APOD) in the neurons (n = 5). (F) Western blot showed the protein expression level of ATF3 in the trigeminal ganglia. (G) Quantitative analysis of the relative ratio of ATF3 per GAPDH in the blots (n = 3). (H) ELISA showed the NGF level in the nerons (n = 5). ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001. NGF, nerve growth factor; ATF3, activating transcription factor 3; NTF3, neurotrophin-3; NTF5, neurotrophin-5; GDNF, glial cell line-derived neurotrophic factor; FASN, fatty acid synthase; SREBF1, sterol regulatory element-binding transcription factor 1; APOD, apolipoprotein D; H3Q5Ser, histone H3 glutamine 5 serotonylation; H3K4me3, histone H3 lysine 4 trimethylation; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Tuj1, neuron-specific class III β-tubulin.

Article Snippet: The used primary antibodies included H3Q5ser (PTM-1420, PTM Bio, Hangzhou, China), H3K4me3 (PTM-5019, PTM Bio), H3 (PTM-6613, PTM Bio), TGM2 (3557, Cell Signaling Technology, Boston, MA, USA), ATF3 (18665, Cell Signaling Technology), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vitro, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Binding Assay

Journal: Journal of Dental Sciences

Article Title: Serotonin promotes lip sensory recovery after inferior alveolar nerve transection via histone serotonylation

doi: 10.1016/j.jds.2025.04.001

Figure Lengend Snippet: GK921 delayed the lip sensory recovery after inferior alveolar nerve transection by reducing histone serotonylation in vivo . (A) Quantitative sensory testing with Von Frey filaments showed the lower lip sensation after transection alone (Trans group) versus transection plus exogenous serotonin group (administration of 50 μM GK921) (Trans + GK921 group) in mice, measured every four days for 8 weeks (n = 5). (B) Immunofluorescence showed the level of H3Q5Ser, H3K4me3, Tuj in the trigeminal ganglia. Bar indicated 20 μm. (C) Quantitative analysis of immunofluorescence strength of H3Q5Ser and H3K4me3 in Tuj-positive area (n = 5). (D) Western blot showed the level of H3Q5Ser and H3K4me3 in the trigeminal ganglia. (E) Quantitative analysis of the relative ratio of H3Q5Ser per H3K4me3 in the blots (n = 3). (F) Quantitative real-time polymerase chain reaction showed the genes related to axon regeneration (including NGF, ATF3, NTF3, NTF5, and GDNF) and lipid metabolism (FASN, SREBF1, and APOD) in the trigeminal ganglia (n = 5). (G) Western blot showed the protein expression level of ATF3 in the trigeminal ganglia. (H) Quantitative analysis of the relative ratio of ATF3 per GAPDH in the blots (n = 3). (I) ELISA showed the NGF level in the trigeminal ganglia (n = 5). ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001. NGF, nerve growth factor; ATF3, activating transcription factor 3; NTF3, neurotrophin-3; NTF5, neurotrophin-5; GDNF, glial cell line-derived neurotrophic factor; FASN, fatty acid synthase; SREBF1, sterol regulatory element-binding transcription factor 1; APOD, apolipoprotein D; H3Q5Ser, histone H3 glutamine 5 serotonylation; H3K4me3, histone H3 lysine 4 trimethylation; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; Tuj1, neuron-specific class III β-tubulin.

Article Snippet: The used primary antibodies included H3Q5ser (PTM-1420, PTM Bio, Hangzhou, China), H3K4me3 (PTM-5019, PTM Bio), H3 (PTM-6613, PTM Bio), TGM2 (3557, Cell Signaling Technology, Boston, MA, USA), ATF3 (18665, Cell Signaling Technology), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vivo, Immunofluorescence, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Binding Assay

Journal: Frontiers in immunology

Article Title: Hepatocyte-specific mitogen-activated protein kinase phosphatase 1 in sexual dimorphism and susceptibility to alcohol induced liver injury.

doi: 10.3389/fimmu.2024.1316228

Figure Lengend Snippet: FIGURE 6 Increased ER stress in alcohol-fed hepatocyte-specific Mkp1 knockout mice. (A) RT-qPCR analysis of genes associated with ER stress in total liver tissue from pair-fed and alcohol-fed male mice in the chronic plus binge model. (B) Western blot analysis of hepatic ATF3, ATF4 and CHOP proteins in males. (C) hepatic ER stress gene expression in females, (D) Western blot analysis of hepatic ATF3, ATF4 and CHOP proteins in females. Two-way ANOVA, *P ≤0.05, **P ≤0.01, ***P ≤0.001, ****P ≤0.0001 compared to their PF counterparts, aP ≤0.01 compared to AF-f/f.

Article Snippet: Antibodies to GAPDH (#5174), phospho-JNK (Thr183/Tyr185, #9251), JNK (#9252), CHOP (#5554), ATF3 (#18665), ATF4 (#11815), phospho-Akt (Ser473) and Akt (#9272) were purchased from Cell Signaling Technology.

Techniques: Knock-Out, Quantitative RT-PCR, Western Blot, Gene Expression

Journal: Journal of Dental Sciences

Article Title: Fatostatin delayed lip sensory recovery after inferior alveolar nerve transection by inhibiting sterol regulatory element-binding protein 1

doi: 10.1016/j.jds.2025.04.024

Figure Lengend Snippet: Fatostatin impaired lip sensory recovery following inferior alveolar nerve transection in vivo . (A) Quantitative sensory testing using Von Frey filaments showed the sensory recovery of the lower lip in the control and fatostatin-treated groups, measured every 4 days for 40 days. The results show that fatostatin significantly delayed lip sensory recovery compared to the control group. (B) Quantitative real-time PCR analysis of gene expression in the trigeminal ganglia demonstrated that fatostatin treatment significantly decreased the mRNA levels of SREBP1, FASN, ACLY, ATF3, and NGF. (C) Western blot analysis confirmed that fatostatin treatment reduced the expression levels of SREBP1, ACLY, and ATF3 in the trigeminal ganglia compared to the control group. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001. SREBP1, sterol regulatory element-binding protein 1; FASN, fatty acid synthase; ACLY, ATP citrate lyase; ATF3, activating transcription factor 3; NGF, nerve growth factor.

Article Snippet: The used primary antibodies included SREBP1 (sc-13551, Santa Cruz, Dallas, TX, USA), ACLY (sc-517267, Santa Cruz), ATF3 (18665, Cell Signaling Technology, Boston, MA, USA), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vivo, Control, Real-time Polymerase Chain Reaction, Gene Expression, Western Blot, Expressing, Binding Assay

Journal: Journal of Dental Sciences

Article Title: Fatostatin delayed lip sensory recovery after inferior alveolar nerve transection by inhibiting sterol regulatory element-binding protein 1

doi: 10.1016/j.jds.2025.04.024

Figure Lengend Snippet: Fatostatin inhibits axonal growth of primary trigeminal neurons in vitro . (A) Images of cultured primary neurons stained with NF200. The upper panels show the control group (left) and the fatostatin-treated group (right) at 20 × magnification. The lower panels show higher magnification (100 × ) of neurons in both groups. Scale bars in the upper panels indicate 100 μm, and in the lower panels, 20 μm. (B) Quantitative analysis of axonal length in the top 50 neurons, showing that fatostatin significantly reduced axonal growth compared to the control group. (C) Quantitative PCR analysis of key genes related to lipid metabolism and axonal regeneration, including SREBP1, FASN, ACLY, ATF3, and NGF. The results show that fatostatin treatment significantly reduced the expression levels of these genes, with SREBP1 and FASN showing the most prominent changes. (D) Western blot analysis of SREBP1, ACLY, and ATF3 protein expression levels, demonstrating that fatostatin treatment decreased SREBP1, ACLY, and ATF3 protein levels compared to control. GAPDH was used as the loading control. ∗∗∗, P < 0.001, ∗∗∗∗, P < 0.0001. SREBP1, sterol regulatory element-binding protein 1; FASN, fatty acid synthase; ACLY, ATP citrate lyase; ATF3, activating transcription factor 3; NGF, nerve growth factor.

Article Snippet: The used primary antibodies included SREBP1 (sc-13551, Santa Cruz, Dallas, TX, USA), ACLY (sc-517267, Santa Cruz), ATF3 (18665, Cell Signaling Technology, Boston, MA, USA), GAPDH (GB15002, Servicebio, Wuhan, China) After washing in TBST, membranes were incubated with the secondary antibody (ZSGB-BIO, Beijing, China), washed again in TBST, and visualized.

Techniques: In Vitro, Cell Culture, Staining, Control, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Binding Assay