Journal: PLOS One

Article Title: Semaglutide ameliorates diabetes-associated cognitive dysfunction in mouse model of type 2 diabetes

doi: 10.1371/journal.pone.0326897

Figure Lengend Snippet: (A) Hematoxylin and eosin (H&E) staining of hippocampal CA1 region shows preserved neuronal structure in the control, semaglutide and dapagliflozin groups compared to the T2DM group. Black arrows indicate nuclear pyknosis (condensed, shrunken nuclei) in the T2DM group, suggesting neuronal damage. (B) Nissl staining further confirms the preservation of neuronal integrity in control and treated groups, with the T2DM group showing reduced Nissl substance. Black arrows indicate areas of neuronal loss and structural disruption in the hippocampal CA1 region. (C) Immunohistochemical staining for LRP1 (Low-density lipoprotein receptor-related protein 1) reveals reduced expression in the T2DM group, with restoration observed in both semaglutide and dapagliflozin-treated groups. (D) Quantitative analysis of LRP1 expression shows significantly decreased levels in the T2DM group compared to control (*** p < 0.001), while both semaglutide and da p agliflozin treatment significantly restored LRP1 expression compared to the T2DM group (*** p < 0.001). (E) Double immunofluorescence staining for NeuN (neuronal marker, red) and TUNEL (a p optosis marker, green), with DAPI nuclear counterstain (blue). The merged images show substantially increased neuronal apoptosis in the T2DM group compared to control, with both semaglutide and dapagliflozin treatments reducing apoptotic signals. (F) Quantitative analysis of NeuN-positive cells demonstrates significant neuronal loss in the T2DM group compared to control (*** p < 0.001), with both treatments p reserving neuronal populations compared to the untreated T2DM group (*** p < 0.001). (G) Quantitative analysis of TUNEL- p ositive cells confirms significantly increased apoptosis in the T2DM group compared to control (*** p < 0.001), with both semaglutide and da p agliflozin treatments significantly reducing neuronal apoptosis (*** p < 0.001). Data are p resented as mean ± standard deviation (SD).

Article Snippet: Tissue sections were first incubated with primary antibodies against LRP1 (Rabbit, Abclonal, A14439, diluted at 1:200) and the neuronal marker NeuN (Rabbit, Affinity, DF6145, diluted at 1:200), followed by incubation with appropriate secondary antibodies: Goat Anti-Rabbit IgG Secondary Antibody (HRP, sino biological, SSA004, diluted at 1:1000) and Anti-rabbit IgG (H + L), F(ab’)2 Fragment (Alexa Fluor® 555 Conjugate, CST, #44413, diluted at 1:1000).

Techniques: Staining, Control, Preserving, Disruption, Immunohistochemical staining, Expressing, Double Immunofluorescence Staining, Marker, TUNEL Assay, Standard Deviation

Journal: Biology Direct

Article Title: MLL1 promotes placental trophoblast ferroptosis and aggravates preeclampsia symptoms through epigenetic regulation of RBM15/TRIM72/ADAM9 axis

doi: 10.1186/s13062-024-00572-0

Figure Lengend Snippet: Inhibition of MLL1 expression ameliorates symptoms of PE mice and alleviates ferroptosis in placental tissues. A murine model of PE was established by injection of sFlt-1 adenovirus via tail vein at E8.5, and the control mice were injected with adenovirus encoding Fc protein, followed by injection of sh-MLL1 adenovirus, with sh-NC adenovirus as the control. A : Measurement of blood pressure in mice, n = 12. B : Detection of proteinuria in urine, n = 12. C : Weight of placenta and fetus, n = 12. D : Detection of MLL1 expression in placenta by qRT-PCR, n = 6. E : Observation on the pathological structure of placenta and kidney by H&E staining and PAS staining, respectively. The black arrow indicates the placental villous vascular interstitium; ▲ indicates trophoblasts; the green arrows indicates pathological features of glomeruli in renal tissues, n = 6. F-I : Detection of ROS, MDA, Fe 2+ , and GSH levels in placenta, n = 6. J: Detection of MLL1, ASCL4, GPX4, and FTH1 expressions in placenta by Western blot, n = 6. Data in panels BCDFGHI were analyzed by one-way ANOVA, and data in panels AJ were analyzed by two-way ANOVA, followed by Tukey’s multiple comparisons test. ** P < 0.01

Article Snippet: Then, the membranes were detected with the primary antibodies anti-MLL1 (1:2000, NB600-256, Novus Biological Inc., Littleton, CO, USA), RBM15 (1:1000, ab315456, Abcam, Cambridge, MA, USA), YTHDF2 (1:1000, ab220163, Abcam), TRIM72 (1:1000, ab220163, Abcam), ADAM9 (1:10000, ab226459, Abcam), ASCL4 (1:10000, NBP2-92741, NOVUS), GPX4 (1:1000, ab125066, Abcam), and FTH1 (1:1000, 701934, Invitrogen) in tris-buffered saline with Tween 20 (TBS-T) containing 2% (w/v) bovine serum albumin (RPN4201, GE-Healthcare, USA) overnight at 4 °C.

Techniques: Inhibition, Expressing, Injection, Control, Quantitative RT-PCR, Staining, Western Blot

Journal: Biology Direct

Article Title: MLL1 promotes placental trophoblast ferroptosis and aggravates preeclampsia symptoms through epigenetic regulation of RBM15/TRIM72/ADAM9 axis

doi: 10.1186/s13062-024-00572-0

Figure Lengend Snippet: Inhibition of MLL1 promotes the invasion and migration of HTR-8/SVneo cells and represses ferroptosis. HTR-8/SVneo cells were transfected with sh-MLL1, with sh-NC as the control. A : Detection of transfection efficiency by qRT-PCR; then HTR-8/SVneo cells were treated with 10 µmol/L Erastin for 24 h to establish a cell model. B : Detection of MLL1 expression in cells by qRT-PCR. C : Detection of cell viability by CCK-8 assay. D-F : Detection of MDA, Fe 2+ , and GSH levels in cells by kits. G : Detection of ROS levels by fluorescence. H : Detection of MLL1, ASCL4, GPX4, and FTH1 expressions in cells by Western blot. I : Detection of cell invasion and migration by Transwell. The cell experiments were repeated three times independently. Data in panel A were analyzed by t test. Data in panels BCDEFGI were analyzed by one-way ANOVA, and data in panel H were analyzed by two-way ANOVA, followed by Tukey’s multiple comparisons test. ** P < 0.01

Article Snippet: Then, the membranes were detected with the primary antibodies anti-MLL1 (1:2000, NB600-256, Novus Biological Inc., Littleton, CO, USA), RBM15 (1:1000, ab315456, Abcam, Cambridge, MA, USA), YTHDF2 (1:1000, ab220163, Abcam), TRIM72 (1:1000, ab220163, Abcam), ADAM9 (1:10000, ab226459, Abcam), ASCL4 (1:10000, NBP2-92741, NOVUS), GPX4 (1:1000, ab125066, Abcam), and FTH1 (1:1000, 701934, Invitrogen) in tris-buffered saline with Tween 20 (TBS-T) containing 2% (w/v) bovine serum albumin (RPN4201, GE-Healthcare, USA) overnight at 4 °C.

Techniques: Inhibition, Migration, Transfection, Control, Quantitative RT-PCR, Expressing, CCK-8 Assay, Fluorescence, Western Blot

Journal: Biology Direct

Article Title: MLL1 promotes placental trophoblast ferroptosis and aggravates preeclampsia symptoms through epigenetic regulation of RBM15/TRIM72/ADAM9 axis

doi: 10.1186/s13062-024-00572-0

Figure Lengend Snippet: Overexpression of RBM15 partially reverses the inhibitory effect of MLL1 inhibition on ferroptosis of HTR-8/SVneo cells. HTR-8/SVneo cells were transfected with oe-RBM15, with oe-NC as the control. A : Detection of transfection efficiency by qRT-PCR; then HTR-8/SVneo cells were treated with sh-MLL1 for a combined experiment. B : Detection of RBM15, TRIM72, ASCL4, GPX4, and FTH1 in cells by Western blot. C : Detection of cell viability by CCK-8 assay. D-F : Detection of MDA, Fe 2+ , and GSH levels in cells by kits. G : Detection of ROS levels in cells by fluorescence. H : Detection of cell invasion and migration by Transwell. The cell experiments were repeated three times independently. Data in panel A were analyzed by t test. Data in panels CDEFGH were analyzed by one-way ANOVA, and data in panel B were analyzed by two-way ANOVA, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01

Article Snippet: Then, the membranes were detected with the primary antibodies anti-MLL1 (1:2000, NB600-256, Novus Biological Inc., Littleton, CO, USA), RBM15 (1:1000, ab315456, Abcam, Cambridge, MA, USA), YTHDF2 (1:1000, ab220163, Abcam), TRIM72 (1:1000, ab220163, Abcam), ADAM9 (1:10000, ab226459, Abcam), ASCL4 (1:10000, NBP2-92741, NOVUS), GPX4 (1:1000, ab125066, Abcam), and FTH1 (1:1000, 701934, Invitrogen) in tris-buffered saline with Tween 20 (TBS-T) containing 2% (w/v) bovine serum albumin (RPN4201, GE-Healthcare, USA) overnight at 4 °C.

Techniques: Over Expression, Inhibition, Transfection, Control, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Fluorescence, Migration

Journal: Biology Direct

Article Title: MLL1 promotes placental trophoblast ferroptosis and aggravates preeclampsia symptoms through epigenetic regulation of RBM15/TRIM72/ADAM9 axis

doi: 10.1186/s13062-024-00572-0

Figure Lengend Snippet: Inhibition of TRIM72 partially reverses the inhibitory effect of MLL1 inhibition on ferroptosis of HTR-8/SVneo cells. HTR-8/SVneo cells were transfected with sh-TRIM72, with sh-NC as the control. A : Detection of transfection efficiency by qRT-PCR; then HTR-8/SVneo cells were treated with sh-MLL1 for a combined experiment. B : Detection of TRIM72, ASCL4, GPX4, and FTH1 expressions in cells by Western blot. C : Detection of cell viability by CCK-8 assay. D-F : Detection of MDA, Fe 2+ , and GSH levels in cells by kits. G : Detection of ROS levels in cells by fluorescence. H : Detection of cell invasion and migration by Transwell. The cell experiments were repeated three times independently. Data in panel A were analyzed by t test. Data in panels CDEFGH were analyzed by one-way ANOVA, and data in panel B were analyzed by two-way ANOVA, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01

Article Snippet: Then, the membranes were detected with the primary antibodies anti-MLL1 (1:2000, NB600-256, Novus Biological Inc., Littleton, CO, USA), RBM15 (1:1000, ab315456, Abcam, Cambridge, MA, USA), YTHDF2 (1:1000, ab220163, Abcam), TRIM72 (1:1000, ab220163, Abcam), ADAM9 (1:10000, ab226459, Abcam), ASCL4 (1:10000, NBP2-92741, NOVUS), GPX4 (1:1000, ab125066, Abcam), and FTH1 (1:1000, 701934, Invitrogen) in tris-buffered saline with Tween 20 (TBS-T) containing 2% (w/v) bovine serum albumin (RPN4201, GE-Healthcare, USA) overnight at 4 °C.

Techniques: Inhibition, Transfection, Control, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Fluorescence, Migration

Journal: Biology Direct

Article Title: MLL1 promotes placental trophoblast ferroptosis and aggravates preeclampsia symptoms through epigenetic regulation of RBM15/TRIM72/ADAM9 axis

doi: 10.1186/s13062-024-00572-0

Figure Lengend Snippet: Overexpression of ADAM9 partially reverses the inhibitory effect of MLL1 inhibition on ferroptosis of HTR-8/SVneo cells. HTR-8/SVneo cells were transfected with oe-ADAM9, with oe-NC as the control. A : Detection of transfection efficiency by qRT-PCR; then HTR-8/SVneo cells were treated with sh-MLL1 for a combined experiment. B : Detection of ADAM9, ASCL4, GPX4, and FTH1 expressions in cells by Western blot. C : Detection of cell viability by CCK-8 assay. D-F : Detection of MDA, Fe 2+ , and GSH levels in cells by kits. G : Detection of ROS levels in cells by fluorescence. H: Detection of cell invasion and migration by Transwell. The cell experiments were repeated three times independently. Data in panel A were analyzed by t test. Data in panels CDEFGH were analyzed by one-way ANOVA, and data in panel B were analyzed by two-way ANOVA, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01

Article Snippet: Then, the membranes were detected with the primary antibodies anti-MLL1 (1:2000, NB600-256, Novus Biological Inc., Littleton, CO, USA), RBM15 (1:1000, ab315456, Abcam, Cambridge, MA, USA), YTHDF2 (1:1000, ab220163, Abcam), TRIM72 (1:1000, ab220163, Abcam), ADAM9 (1:10000, ab226459, Abcam), ASCL4 (1:10000, NBP2-92741, NOVUS), GPX4 (1:1000, ab125066, Abcam), and FTH1 (1:1000, 701934, Invitrogen) in tris-buffered saline with Tween 20 (TBS-T) containing 2% (w/v) bovine serum albumin (RPN4201, GE-Healthcare, USA) overnight at 4 °C.

Techniques: Over Expression, Inhibition, Transfection, Control, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Fluorescence, Migration

Journal: Biology Direct

Article Title: MLL1 promotes placental trophoblast ferroptosis and aggravates preeclampsia symptoms through epigenetic regulation of RBM15/TRIM72/ADAM9 axis

doi: 10.1186/s13062-024-00572-0

Figure Lengend Snippet: Overexpression of RBM15 partially reversed the ameliorative effect of MLL1 inhibition on symptoms in PE mice. A murine model of PE was established by injection of sFlt-1 adenovirus via tail vein at E8.5, and the control mice were injected with adenovirus encoding Fc protein, followed by injection of oe-RBM15 adenovirus for a combined experiment with sh-MLL1, with oe-NC adenovirus as the control. A : Detection of RBM15 expression in placenta by qRT-PCR. B : Measurement of blood pressure in mice, n = 12. C : Detection of proteinuria in urine, n = 12. D : Weight of placenta and fetus, n = 12. E : Observation on the pathological structure of placenta and kidney by H&E staining and PAS staining, respectively. The black arrow indicates the placental villous vascular interstitium; ▲ indicates trophoblasts; the green arrows indicates pathological features of glomeruli in renal tissues, n = 6. F-I : Detection of ROS, MDA, Fe 2+ , and GSH levels in placenta, n = 6. J : Detection of RBM15, TRIM72, ADAM9, ASCL4, GPX4, and FTH1 expressions in placenta by Western blot, n = 6. Data in panels ACDFGHI were analyzed by one-way ANOVA, and data in panels BJ were analyzed by two-way ANOVA, followed by Tukey’s multiple comparisons test. * P < 0.05, ** P < 0.01

Article Snippet: Then, the membranes were detected with the primary antibodies anti-MLL1 (1:2000, NB600-256, Novus Biological Inc., Littleton, CO, USA), RBM15 (1:1000, ab315456, Abcam, Cambridge, MA, USA), YTHDF2 (1:1000, ab220163, Abcam), TRIM72 (1:1000, ab220163, Abcam), ADAM9 (1:10000, ab226459, Abcam), ASCL4 (1:10000, NBP2-92741, NOVUS), GPX4 (1:1000, ab125066, Abcam), and FTH1 (1:1000, 701934, Invitrogen) in tris-buffered saline with Tween 20 (TBS-T) containing 2% (w/v) bovine serum albumin (RPN4201, GE-Healthcare, USA) overnight at 4 °C.

Techniques: Over Expression, Inhibition, Injection, Control, Expressing, Quantitative RT-PCR, Staining, Western Blot