Journal: Stem cells (Dayton, Ohio)

Article Title: β-Arrestin 2 and Epac2 Cooperatively Mediate DRD1-Stimulated Proliferation of Human Neural Stem Cells and Growth of Human Cerebral Organoids.

doi: 10.1093/stmcls/sxac046

Figure Lengend Snippet: Figure 3. p38 MAPK functioned as the downstream of β-arrestin 2. (A) Protein levels of phosphorylated p38 MAPK in WT and β-arrestin 2 KO cells after SKF83566 treatment in 3L NSC. (B) Statistic of protein bands in different times. (C) Protein levels of phosphorylated JNK in WT and β-arrestin 2 KO cells after SKF83566 treatment. (D) Statistic of protein bands in different times. (E) Protein levels of phosphorylated ERK1/2 in WT and β-arrestin 2 KO cells after SKF83566 treatment. (F) Statistic of protein bands in different times. (G, H) The effect of SKF83566 on human NSCs proliferation in the presence of p38 MAPK inhibitors (SB203580, SKF86002). (I) The effect of SKF83566 on human NSCs proliferation in the presence of JNK inhibitor SP600125. (J) The effect of SKF83566 on human NSCs proliferation in the presence of ERK inhibitor PD98059. (K, L) The number of secondary neurospheres formed from SKF83566-treated control or p38 MAPK-knockdown NSCs. For B, D, F-J, N = 3 independent experiments. Data are mean ± SEM. ***P < .001, **P < .01, *P < .05, 2-way ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: SKF83566 was purchased from Tocris Bioscience; ESI-05, HJC0350, SB203580, SKF86002, PD98059, SP600125 were purchased from MedChemExpress; Cell CountingLite 2.0 Luminescent cell viability assay was purchased from Vazyme.

Techniques: Control, Knockdown

Journal: Cell Death Discovery

Article Title: Unraveling ferroptosis in osteogenic lineages: implications for dysregulated bone remodeling during periodontitis progression

doi: 10.1038/s41420-024-01969-6

Figure Lengend Snippet: A Schematic illustration of drug administration during ligature. B Representative 3D micro-CT images of murine upper molars and alveolar bone in the vehicle, Lip-1, and erastin groups. Scale bar, 1 mm. C Quantification of the distance between the cementum enamel junction and alveolar bone crest (CEJ-ABC distance). D The bone volume over total volume (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th), and trabecular spacing (Tb.Sp) of the ROIs in the maxillary alveolar bone on the ligature side. E Representative images of H&E-stained sagittal sections of the upper first molars on the ligature side and magnified views of the gingiva and alveolar bone distal to the upper first molar. The green arrows indicate inflammatory cells. The blue arrows indicate empty osteocyte lacunae. Scale bar, 200 μm (panoramic view)/50 μm (magnified view). F Representative TUNEL staining of dead cells in distal regions of upper first molars. Magnified views of the gingiva or alveolar bone are shown in the yellow or orange frames, respectively. Scale bar, 100 μm (panoramic view)/10 μm (magnified view). G Representative immunofluorescence images of 4-HNE, GPX4, and TfR1 expression in distal regions of the upper first molars. Scale bar, 100 μm. H The percentage of TUNEL-positive cells was quantified. I Quantification of the mean fluorescence intensity of 4-HNE, GPX4, and TfR1 in the alveolar bone of the vehicle, Lip-1, and erastin groups. G gingiva, AB alveolar bone, P periodontal ligament, D dentin, DP dental pulp. The data are shown as the means ± SDs; n = 3 mice; ns not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Once the ligature was placed (Day 0), 10 mg/kg Lip-1 (T2367, TargetMOI), 30 mg/kg Erastin (T1765, TargetMOI), or an equal volume of vehicle was injected intraperitoneally once every other day over the 10-day period.

Techniques: Micro-CT, Staining, TUNEL Assay, Immunofluorescence, Expressing, Fluorescence

Journal: Cell Death Discovery

Article Title: Unraveling ferroptosis in osteogenic lineages: implications for dysregulated bone remodeling during periodontitis progression

doi: 10.1038/s41420-024-01969-6

Figure Lengend Snippet: A Representative immunofluorescence images showing RANKL expression in distal regions of upper first molars in the vehicle, Lip-1, and erastin groups. The lower panel shows magnified views of the boxed areas in the upper panel. Scale bar, 50 μm. B Quantification of the mean fluorescence intensity of RANKL in the alveolar bone. C Representative immunofluorescence images of RUNX2 expression in distal regions of upper first molars. The lower panel shows magnified views of the boxed areas in the upper panel. Scale bar, 50 μm (panoramic view)/10 μm (magnified view). D Quantification of the mean fluorescence intensity of RUNX2 in alveolar bone. E Representative images of TRAP staining in the vehicle, Lip-1, and erastin groups. Magnified views of alveolar bone in the interradicular region or distal alveolar bone are shown in the blue or red frames, respectively. The white arrows indicate TRAP-positive cells. Scale bar, 200 μm (panoramic view)/40 μm (magnified view). F Quantification of TRAP-positive cells per mm of bone surface. G gingiva, AB alveolar bone. The data are shown as the means ± SDs; n = 3 mice; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Once the ligature was placed (Day 0), 10 mg/kg Lip-1 (T2367, TargetMOI), 30 mg/kg Erastin (T1765, TargetMOI), or an equal volume of vehicle was injected intraperitoneally once every other day over the 10-day period.

Techniques: Immunofluorescence, Expressing, Fluorescence, Staining

Journal: Cell Death Discovery

Article Title: Unraveling ferroptosis in osteogenic lineages: implications for dysregulated bone remodeling during periodontitis progression

doi: 10.1038/s41420-024-01969-6

Figure Lengend Snippet: A TUNEL assay showing dead MLO-Y4 cells after treatment with vehicle, LPS, or LPS + Lip-1. Scale bar, 50 μm (panoramic view)/10 μm (magnified view). B Statistics of the percentage of TUNEL-positive osteocytes. C CCK-8 assay of treated MLO-Y4 cells. D C11-BODIPY staining showing nonoxidized lipids (red) and oxidized lipids (green) in treated MLO-Y4 cells. Scale bar, 10 μm. E Quantification of the mean fluorescence intensity of C11-BODIPY (green/red ratio). F Measurement of intracellular ROS levels in treated MLO-Y4 cells. G Relative mRNA expression of Slc7a11 and Ncoa4 in treated MLO-Y4 cells. H – K Western blot analysis of TfR1, RANKL, OPG, GPX4, and IL-6 expression in treated MLO-Y4 cells. L Quantification of the relative RANKL and OPG protein expression and the RANKL/OPG ratio. The data are shown as the means ± SDs; n = 3 biological replicates; ns not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Once the ligature was placed (Day 0), 10 mg/kg Lip-1 (T2367, TargetMOI), 30 mg/kg Erastin (T1765, TargetMOI), or an equal volume of vehicle was injected intraperitoneally once every other day over the 10-day period.

Techniques: TUNEL Assay, CCK-8 Assay, Staining, Fluorescence, Expressing, Western Blot