Journal: The Journal of Clinical Investigation

Article Title: Aryl hydrocarbon receptor sulfenylation promotes glycogenolysis and rescues cancer chemoresistance

doi: 10.1172/JCI170753

Figure Lengend Snippet: ( A ) Immunoblot of immunoprecipitations of STBD1 or HSP90 in lysates from MCF-7 or MCF-7/DDP cells treated with DDP for 24 hours. ( B ) Strategy for detecting sulfenic acid modification of AHR with dimedone. LC-MS/MS analysis of dimedone-labeled AHR. Analysis of the y-ions indicates the formation of dimedone adduct (+138.07 Da). ( C ) MCF-7/DDP or A549/5-Fu cells were treated with DDP or 5-Fu for 24 hours. The location of AHR (green), dimedone (red), and STBD1 (cyan) was observed under super-resolution microscope. The arrows indicate the colocation of the sulfenylated AHR and STBD1. Scale bars, 10 μm. ( D ) Separated glycogen-enriched pellets from MCF-7/DDP or A549/5-Fu cells treated with DDP or 5-Fu for 24 hours were resuspended in IP buffer, and immunoblot with dimedone of immunoprecipitations of AHR was analyzed. ( E ) Purified Flag-tagged (Flag-AHR) protein was treated with different doses of H 2 O 2 for 30 minutes. Sulfenic acid modification of dimedone-labeled AHR was analyzed by Western blot. ( F ) Purified Flag-AHR protein was treated with different doses of H 2 O 2 for 30 minutes. The binding between modified AHR and HSP90 was measured by biolayer interferometry (BLI). ( G ) HEK293T cells transfected with the indicated combinations of Flag-HSP90, Myc-AHR, or Myc-AHR(C300A) were treated with H 2 O 2 for 30 minutes. Immunoprecipitations of Flag or STBD1 were analyzed by Western blot.

Article Snippet: The recombinant human AHR protein was purified according to the method described in the former context, while the HSP90 protein (active) was purchased from Abcam.

Techniques: Western Blot, Modification, Liquid Chromatography with Mass Spectroscopy, Labeling, Super-Resolution Microscopy, Purification, Binding Assay, Transfection

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: SOX2 + cells are distributed in the optic tectum and thalamus. ( A ) Representative images of a tadpole (left) and whole-mount immunofluorescent staining with an anti-SOX2 antibody (right) at stage 49 Xenopus . The red square indicates the whole optic tectum and the thalamus. Scale bar: 100 μm. ( B ) Representative images showing the colabeling with SOX2 and Nkx2.2 in the brain. The dotted white line indicates the outline of the optic tectum (OT). The white line represents the boundary of the SOX2 and Nkx2.2 immunoreactive thalamus (Th). Scale bar: 50 μm. ( C ) Six representative coronal planes of the whole brain with SOX2 immunostaining are shown at stage 49 Xenopus ( Ca – Cf ). Scale bar: 20 μm. ( D ) One representative sagittal section was immunostained with an anti-SOX2 antibody. The white lines (a–e) depict the positions of coronal sections for ( Ca – Ce ). Scale bar: 100 μm. III: third ventricle; IV: fourth ventricle; A: anterior; ABB: alar basal boundary; BH: basal hypothalamus; c-Th: caudal thalamus; D: dorsal; Hb: habenula; M: middle ventricle; Mes: mesencephalon; oc: optic chiasm; P: posterior; p1–3: prosomere1–3; Pa: pallium; PTh: prethalamus; r1–r7: rhomeres1–7; Spa: subpallium; OT: optic tectum; R: rostral; r-Th: rostral thalamus; Th: thalamus; Zli: zona limitans intrathalamica.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques: Staining, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: The majority of thalamic SOX2 + cells are HuC/D + or tubulin + neurons. ( A ) Colabeling of SOX2 and BrdU showing that only a few SOX2 + cells ( Aa ) are BrdU + cells ( Ab ) in the thalamus. Arrowheads indicate the SOX2 − and BrdU + cells ( Ac ). Arrows indicate the SOX2 + and BrdU + cells ( Ac ). Scale bar: 20 μm. ( B ) Coimmunostaining of BrdU and PCNA in the thalamus. Arrows indicate the BrdU + and PCNA + cells. Scale bar: 20 μm. ( C ) Coimmunostaining of SOX2 ( Ca ) and PCNA ( Cb ) in the thalamus. Arrowheads indicate the SOX2 + and PCNA − cells. Arrows indicate the SOX2 + and PCNA + neurons ( Cc , Cd ). Scale bar: 20 μm. ( D ) Coimmunostaining of SOX2 ( Da ) and HuC/D ( Db ) in the thalamus. Arrows indicate the SOX2 + and HuC/D + neurons ( Dc , Dd ). Scale bar: 20 μm. ( E ) Colabeling of SOX2 ( Ea ) and tubulin ( Eb ) in the thalamus. Arrows indicate the SOX2 + and tubulin + neurons ( Ec , Ed ). Scale bar: 20 μm. ( F ) Thalamic cells were immunostained with an anti-SOX2 antibody (red, Fb ) followed by transfection with pSOX2::GFP (green, Fa ), showing that the SOX2 + cells exhibited neuronal morphology with predicted dendrites (arrowheads, Fc , Fd ). Arrows indicate the GFP–expressing and SOX2–immunoreactive cells. Scale bar: 20 μm.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques: Transfection, Expressing

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: Visual deprivation changes the balance between proliferation and differentiation in the thalamus. ( A , B ) Representative fluorescent images showing BrdU– ( Ab ) and SOX2–labeled ( Aa ) cells in Ctrl ( Aa – Ac ) and VD ( Ba – Bc ) thalamus. The white square indicates the BrdU– and SOX2–labeled cells in the zoomed–in thalamus ( Aa – Ac ). White dotted lines indicate the boundary of the thalamus ( Ac ). Arrows indicate the SOX2 + and BrdU + cells ( Ad – Af , Bd – Bf ). Scale bar: 50 μm. Zoom Scale bar: 20 μm. ( C – E ) Summary data show that VD increased BrdU + cells ( C ), decreased SOX2 + cells ( D ), and increased SOX2 + /BrdU + cells ( E ). N = 6, 6 for Ctrl and VD. ( F ) A representative immunofluorescent image showing z–stack for SOX2– and BrdU–labeled cells. Scale bar: 20 μm. ( G , H ) Summary data showing that VD increased BrdU + cells ( G ) and decreased SOX2 + cells ( H ) in VD–treated tadpoles. N = 4, 6 for Ctrl and VD. ( I , J ) Representative immunofluorescent images showing SOX2– and HuC/D–labeled cells in the thalamus of Ctrl ( Ia – Id ) and VD ( Ja – Jd ) tadpoles. The white square indicates the zoomed–in images. Arrows indicate the SOX2 + and HuC/D + cells ( Ie – Ih , Je – Jh ). Scale bar: 50 μm. Zoom scale bar: 20 μm. ( K – M ) Summary of data showing that VD decreased HuC/D + , SOX2 + , and HuC/D + /SOX2 + cells. N = 5, 8 for Ctrl and VD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques: Labeling

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: Visual deprivation reduces nuclearized β-catenin and differentiated neurons in the thalamus. ( A , B ) Representative fluorescent images show β-catenin– and SOX2–labeled cells in the Ctrl ( Aa – Ad ) and VD ( Ba – Bd ) thalamus. The white square indicates the β-catenin– and SOX2–labeled cells in the zoom of the thalamus. Arrows indicate the expressions of β-catenin– and SOX2 in the nuclei. Arrowheads indicate that β-catenin was expressed in the cytoplasm ( Ae – Ah , Be – Bh ). Scale bar: 50 μm, zoom scale bar: 20 μm. ( C , D ) Summary data show that VD decreased β-catenin nuclear localization in labeled cells. N = 5, 6 for β-catenin and SOX2. ( E ) Western blot analysis of homogenates from Ctrl and VD–treated brains using the anti-SOX2, anti-β-catenin, or anti-Phospho-β-catenin (P-β-Cat) antibody. ( F – H ) Summary of data showing the relative intensities of SOX2 (( F ), N = 7), β-catenin (( G ), N = 10), and P-β-Cat (( H ), N = 13) to GAPDH. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques: Labeling, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: SOX2-MO and β-Cat-MO knockdown decreased β-catenin and SOX2 expression. ( A ) Western blot analysis of homogenates from Ctrl-MO–, SOX2-MO– and β-Cat-MO–transfected brains using the anti-SOX2 or anti-β-catenin antibody. ( B ) Quantification results show that SOX2 expression was significantly decreased in SOX2-MO or β-Cat-MO expressing cells compared to Ctrl-MO expressing cells. N = 4. ( C ) Summary of data showing that the relative intensity of the β-catenin group was significantly decreased compared to that of the Ctrl-MO group. N = 12. ( D ) Representative fluorescent images showing the immunostaining of β-catenin and BrdU in the Ctrl-MO– ( Da – Dc ) or β-Cat-MO–transfected ( Dd – Df ) cells. Scale bar: 20 μm. ( E ) Summary data show that the knockdown of β-catenin increased the number of BrdU + cells. N = 7, 7 for Ctrl-MO and β-Cat-MO. ( F ) Representative images ( Fa – Ff ) showing the immunostaining of β-catenin ( Fa , Fd ) and HuC/D ( Fb , Fe ). Scale bar: 20 μm. ( G ) Summary data show that the knockdown of β-catenin decreased the number of HuC/D + cells. N = 8, 4 for Ctrl-MO and β-Cat-MO. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques: Knockdown, Expressing, Western Blot, Transfection, Immunostaining

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: The visual deprivation-induced decrease in SOX2 + and β-catenin + cells was prevented by TDZD-8. ( A , B ) Western blot analysis showing that the relative intensity of β-catenin to GAPDH was significantly decreased by IWR-1-endo but increased by TDZD-8. N = 4. ( C , D ) Western blot analysis showing that VD–induced decrease in β-catenin expression was blocked by TDZD-8 treatment. N = 8. ( E , F ) VD–induced increase in P-β-Cat was prevented by TDZD-8 treatment. N = 5. ( G ) Representative immunofluorescent images showing SOX2– and β-catenin–labeled cells in the thalamus of Ctrl ( Ga – Gd ), VD ( Ge – Gh ), TDZD-8 ( Gi – Gl ), and VD + TDZD-8 ( Gm – Gp ) tadpoles. Arrows indicate the double–labeling cells in the thalamus. Scale bar: 20 μm. ( H , I ) Summary of data showing that VD decreased SOX2 + ( H ) and β-catenin + ( I ) cells. TDZD-8 prevented the VD–induced decrease in SOX2 + and β-catenin + cells in the thalamus. N = 7, 4, 5, 5 for Ctrl, VD, TDZD-8, VD + TDZD-8. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques: Western Blot, Expressing, Labeling

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: SOX2 is selectively associated with β-catenin. ( A ) Coimmunoprecipitation of SOX2 with β-catenin. N = 3 experiments. ( B ) Coimmunoprecipitation of β-catenin with SOX2. N = 4 experiments. ( C ) Coimmunoprecipitation of SOX2 with β-catenin in Ctrl and VD–treated thalamus. ( D ) Summary of data showing that VD increases the interaction between SOX2 with β-catenin. N = 6. * p < 0.05. ( E , F ) Coimmunoprecipitation in homogenates from the mouse thalamus. Coimmunoprecipitation of β-catenin with SOX2 ( E ). Coimmunoprecipitation of SOX2 with β-catenin ( F ). N = 2 experiments.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: List of antibodies.

Article Snippet: SOX2, rabbit , Recombinant human SOX2 , Abcam , ab97959 , AB_2341193 , 1:200 (IF).

Techniques: Recombinant

Journal: International Journal of Molecular Sciences

Article Title: β-Catenin and SOX2 Interaction Regulate Visual Experience-Dependent Cell Homeostasis in the Developing Xenopus Thalamus

doi: 10.3390/ijms241713593

Figure Lengend Snippet: List of antibodies.

Article Snippet: SOX9, rabbit , Recombinant human SOX9 , Abcam , ab185230 , AB_2715497 , 1:200 (IF).

Techniques: Recombinant

Journal: JACC: Basic to Translational Science

Article Title: Empagliflozin Suppresses the Differentiation/Maturation of Human Epicardial Preadipocytes and Improves Paracrine Secretome Profile

doi: 10.1016/j.jacbts.2023.05.007

Figure Lengend Snippet: Secreted Protein Levels From the Empagliflozin-Treated Adipocytes Adipocyte-secreted protein levels of (A) IL-6 and (B) MCP-1 significantly decreased in a dose-dependent manner upon incubation with empagliflozin, whereas those of (C) IL-1α, (D) IL-1β, and (E) TGF-β1 did not decrease. Data are presented as mean ± SD and compared using Student’s t -test. ∗ P < 0.05 and ∗∗∗ P < 0.001. Abbreviations as in Figures 2 and .

Article Snippet: iPS-ACM (Axol Bioscience) was incubated with human recombinant IL-6 (1 ng/mL, Abcam) for 2 days.

Techniques: Incubation

Journal: JACC: Basic to Translational Science

Article Title: Empagliflozin Suppresses the Differentiation/Maturation of Human Epicardial Preadipocytes and Improves Paracrine Secretome Profile

doi: 10.1016/j.jacbts.2023.05.007

Figure Lengend Snippet: Clinical Validation of the Impact of IL-6 and MCP-1 Using Data of the Cohort Study (A) Gene expression levels of IL6 in the EAT were significantly attenuated in the diabetic patients treated with SGLT2 inhibitor compared to those treated with other antidiabetic drugs. (B) Those of MCP1 was not significantly affected. (C) IL6 mRNA levels in the EAT were significantly increased in the patients with atrial fibrillation compared to those without atrial fibrillation. (D) No significant difference was observed in MCP1 between the 2 groups. (E) Patients with high IL6 expression in EAT have higher plasma NT-proBNP levels. (F) No significant difference was observed in NT-proBNP levels between the patients with high MCP1 expression in EAT and low MCP1 expression in EAT. Data are presented as mean ± SD and compared using the Mann-Whitney U test. ∗ P < 0.05. NT-proBNP = N-terminal pro–B-type natriuretic peptide; other abbreviations as in Figures 1 , , , and .

Article Snippet: iPS-ACM (Axol Bioscience) was incubated with human recombinant IL-6 (1 ng/mL, Abcam) for 2 days.

Techniques: Expressing, MANN-WHITNEY