Journal: Cells

Article Title: JNK Signalling Regulates Self-Renewal of Proliferative Urine-Derived Renal Progenitor Cells via Inhibition of Ferroptosis

doi: 10.3390/cells12172197

Figure Lengend Snippet: Inhibition of JNK reduces the proliferation of UdRPCs. ( A ) Morphology of the three UdRPCs UM51, UM27 and UF21 with or without AEG3482 treatment after 72 h. Scale bars represent 100 µm. ( B ) Ki67 expression in UdRPCs treated with or not treated with AEG3482 for 72 h. Scale bars represent 50 µm. ( C ) Ki67 proliferation assay for JNK-inhibited UdRPCs (n = 10; * p -value < 0.05, *** p -value < 0.001). ( D ) Bar graph of PI-measured cell death in JNK-inhibited UdRPCs compared to untreated controls (n = 5, * p -value < 0.05, *** p -value < 0.001). Error bars indicate STDEV. ( E ) mRNA expression of nephron progenitor markers SIX2, SALL1, VCAM1 and Ki67. Mean values were normalised to the housekeeping gene RPL37A. ( F ) Gene expression of cell cycle-related genes for the time points 24 h, 72 h and 120 h depicted in a Pearson’s heatmap.

Article Snippet: After JNK inhibition, cells were fixed with 4% paraformaldehyde (Polysciences, Warrington, PA, USA) and stained with the antibody anti-mouse KI67, 1:200 (CST).

Techniques: Inhibition, Expressing, Proliferation Assay

Journal: Cells

Article Title: JNK Signalling Regulates Self-Renewal of Proliferative Urine-Derived Renal Progenitor Cells via Inhibition of Ferroptosis

doi: 10.3390/cells12172197

Figure Lengend Snippet: Inhibition of JNK reduces the proliferation of UdRPCs. ( A ) Morphology of the three UdRPCs UM51, UM27 and UF21 with or without AEG3482 treatment after 72 h. Scale bars represent 100 µm. ( B ) Ki67 expression in UdRPCs treated with or not treated with AEG3482 for 72 h. Scale bars represent 50 µm. ( C ) Ki67 proliferation assay for JNK-inhibited UdRPCs (n = 10; * p -value < 0.05, *** p -value < 0.001). ( D ) Bar graph of PI-measured cell death in JNK-inhibited UdRPCs compared to untreated controls (n = 5, * p -value < 0.05, *** p -value < 0.001). Error bars indicate STDEV. ( E ) mRNA expression of nephron progenitor markers SIX2, SALL1, VCAM1 and Ki67. Mean values were normalised to the housekeeping gene RPL37A. ( F ) Gene expression of cell cycle-related genes for the time points 24 h, 72 h and 120 h depicted in a Pearson’s heatmap.

Article Snippet: Primary antibodies were diluted as follows: anti-mouse KI67 (1:200), anti-rabbit cJUN (1:400) (CST), anti-rabbit phospho-cJUN (1:800) (CST).

Techniques: Inhibition, Expressing, Proliferation Assay

Journal: Nature Communications

Article Title: Human forebrain organoid-based multi-omics analyses of PCCB as a schizophrenia associated gene linked to GABAergic pathways

doi: 10.1038/s41467-023-40861-2

Figure Lengend Snippet: a Workflow of hFOs culture in this study. Scale bar, 200 μm and 300 μm. b , c RT-qPCR analysis for PCCB expression in hiPSCs ( b ) and hFOs ( c ), n = four technical replicates per group. d Immunostaining characterization for representative control hFOs. N ≥ three biologically independent samples in each group. Cortical plate marker, MAP2; intermediate zone marker, TBR2; ventricular zone markers, SOX2 and ki67; forebrain-specific markers, FOXG1 and PAX6. Scale bar, 50 μm. e PCA plot for the PCCB knockdown and control hFOs. f Volcano plot of DEGs between the PCCB knockdown and control hFOs. Upregulated genes are shown with red dots and downregulated genes are shown with blue dots. g , h GO and KEGG analysis for the PCCB knockdown-induced upregulated ( g ) and downregulated DEGs ( h ), respectively. GO terms are shown with red bars, and KEGG pathways are shown with blue bars. i PPI network analysis for 350 downregulated DEGs shared in both PCCB -G1 and PCCB -G2 hFOs. The top 10 hub nodes are shown with orange nodes. j RT-qPCR analysis for GABRA1 , GABRA2 , GABRB2 , and GABRB3 (The hub nodes in the PPI network), n = four technical replicates per group. Data are shown as Mean ± SD. The unpaired two-tailed t-test was used to assess difference between the PCCB -NC and PCCB -G1 or PCCB -G2 group. ** P < 0.01, *** P < 0.001, **** P < 0.0001. Source data underlying b , c , and j are provided as a Source Data file.

Article Snippet: Primary antibodies used in this study were mouse anti-Ki67 (1:1,000, Cell Signaling Technologies, 9449), rabbit anti-FOXG1 (1:100, Abcam, ab18259), rabbit anti-TBR2 (1:800, Cell Signaling Technologies, 81493), mouse anti-SOX2 (1:200, Invitrogen, MA1-014), rabbit anti-PAX6 (1:100, Proteintech, 12323-1-AP), rabbit anti-MAP2 (1:200, Proteintech, 17490-1-AP), mouse anti-GAD67 (1:200, Abcam, ab26116) and mouse anti-DlX2 (1:200, Santa cruz, sc-393879).

Techniques: Quantitative RT-PCR, Expressing, Immunostaining, Marker, Two Tailed Test

Journal: iScience

Article Title: Tat-P combined with GAPR1 releases Beclin1 to promote autophagy and improve Bronchopulmonary dysplasia model

doi: 10.1016/j.isci.2023.107509

Figure Lengend Snippet: Tat-P ameliorates the hyperoxia-induced BPD model in vivo A) The body weight change trend of pups at 1–7 days of Con, Hyp and Hyp+Tat-P groups N = 6. (B) Lung morphology of Con, Hyp and Hyp+Tat-P groups, N = 6. (C) Histopathological changes in the lung by H&E, N = 6, quantification of mean linear intercept (MLI) represents a surrogate of average air space diameter, quantification of mean alveolar number (MAN) represents the average number of alveoli. (D) Representative images of Ki67 immunostaining showing the proliferation (brown staining) in rat lung tissue of Con, Hyp and Hyp + Tat-P group, N = 6. (E) Representative images of TUNEL immunostaining showing the apoptosis (green staining) in rat lung tissue of Con, Hyp and Hyp + Tat-P group, N = 6. (F) Western blot used to detect the protein expression of Bax and Bcl-2 in rat lung tissue of Con, Hyp and Hyp + Tat-P group, N = 6. (G) RT-qPCR detect the mRNA expression of SPC in Con, Hyp and Hyp + Tat-P group, N = 6. (H). Western blot was used to detect the protein expression of SPC in Con, Hyp and Hyp + Tat-P group, N = 6. Unpaired t-test, ∗/#/p＜0.05，∗∗/##/p＜0.01, ∗∗∗/###/p＜0.001, ∗∗∗∗/####/p＜0.0001. Con: control, Hyp: Hyperoxia ∗ VS. Con, # VS. Hyp.

Article Snippet: The slides were then incubated with primary antibodies, including goat anti-mouse Ki67 (CST, 1:100 dilution) and TUNEL and incubated, at room temperature (23°C - 25°C) for 1 h. Subsequently, the slides were incubated with a secondary antibody and stained with diaminodibenzidine according to the instructions provided with the Vectastain kit.

Techniques: In Vivo, Immunostaining, Staining, TUNEL Assay, Western Blot, Expressing, Quantitative RT-PCR

Journal: iScience

Article Title: Tat-P combined with GAPR1 releases Beclin1 to promote autophagy and improve Bronchopulmonary dysplasia model

doi: 10.1016/j.isci.2023.107509

Figure Lengend Snippet:

Article Snippet: The slides were then incubated with primary antibodies, including goat anti-mouse Ki67 (CST, 1:100 dilution) and TUNEL and incubated, at room temperature (23°C - 25°C) for 1 h. Subsequently, the slides were incubated with a secondary antibody and stained with diaminodibenzidine according to the instructions provided with the Vectastain kit.

Techniques: Recombinant, TUNEL Assay, Protease Inhibitor, Magnetic Beads, CCK-8 Assay, BIA-KA, Software