Journal: bioRxiv

Article Title: Revealing the Key Regulators of Cell Identity in the Human Adult Pancreas

doi: 10.1101/2020.09.23.310094

Figure Lengend Snippet: (A) Regulon activity based UMAP of 7393 single cells with the alpha cell population highlighted. (B) Top 10 ranked regulons in non-diabetic alpha cells based on regulon specificity score (RSS). Known cell type specific regulons are colored green. (C) Regulon activity based UMAP colored by the regulon activity of JUND and EGR4 showing the cell type specificity of regulons. (D) lmmunohistochemical evaluation of JUND in healthy human adult pancreata showing nuclear JUND expression in alpha, beta (i) and ductal (ii) cells. Representative images are shown (n = 3 patients), Scale bar: 50 μm. (E) lmmunofluorescence evaluation of JUND, INS, GCG and SST at stage 7 of human iPSC beta cell differentiation. Box indicates a GCG+ cell with nuclear JUND expression. Representative images examined for JUND (green), INS (red), GCG (purple) and DAPI (blue, nuclei counterstaining) are shown (n=3 beta cell differentiations). Scale bars: 50 μm. (F) The percentage of cells with nuclear JUND expression within the INS+, GCG+, polyhormonal and INS+/GCG+ cell populations. Results are shown as the normalized mean±s.d. (n=3 beta cell differentiations). (G) Regulon activity based UMAP of 7393 single cells with the beta cell population highlighted. (H) Top 10 ranked regulons in non-diabetic beta cells based on regulon specificity score (RSS). Known cell type specific regulons are colored green. (I) Regulon activity based UMAP colored by the regulon activity of ZNF705D and ASCL2 showing the cell type specificity of regulons. (J) lmmunohistochemical evaluation of ASCL2 in healthy human adult pancreata showing cytoplasmic ASCL2 expression in mainly beta cells. Representative images examined for INS (green), ASCL2 (red) and DAPI (blue, nuclei counterstaining) are shown (n = 3 patients), Scale bar: 50 μm.

Article Snippet: Immunohistochemistry analyses were carried out largely as described previously ( Demine et al. , 2020 ), using primary antibodies against the following proteins: INS (Dako (Agilent), IR002, Ready to use solution), GCG (Sigma-Aldrich, G2654, 1/1000) and JUND (Atlas Antibodies, HPA063029, 1/100) and the following secondary antibodies (ThermoFisher, A-11075, A-21206, A-31571, 1/500).

Techniques: Activity Assay, Expressing, Cell Differentiation

Journal: Cell reports

Article Title: High-glucose-associated YTHDC1 lactylation reduces the sensitivity of bladder cancer to enfortumab vedotin therapy.

doi: 10.1016/j.celrep.2025.115545

Figure Lengend Snippet: Figure 5. YTHDC1 enhances the mRNA stability of NECTIN4 transcription factor JUND in a m6A-dependent manner (A) The Ominer network tool predicts potential transcription factors for NECTIN4. (B) ChIP-seq from JUND on the NECTIN4 promoter region of the ChIP-Atlas. (C and D) HT1376 and RT112 cells were transfected with the specified shRNA or plasmid for 72 h, and, after puromycin screening, the cells were collected for ChIP-qPCR analysis using immunoglobulin (Ig)G or JUND antibodies.

Article Snippet: For immunohistochemistry (IHC) of the tissuemicroarray (#U100Bl01), the following antibodies were used: YTHDC1 (#29441-1-AP, Proteintech, 1:1500 dilution), NECTIN4 (#67721-1-lg, Proteintech, 1:1500 dilution), and JUND (#AF6200, Affinity Biosciences, 1:1000 dilution).

Techniques: ChIP-sequencing, Transfection, shRNA, Plasmid Preparation, ChIP-qPCR

Journal: Cell reports

Article Title: High-glucose-associated YTHDC1 lactylation reduces the sensitivity of bladder cancer to enfortumab vedotin therapy.

doi: 10.1016/j.celrep.2025.115545

Figure Lengend Snippet: Figure 6. YTHDC1 regulates the sensitivity of BC to EV through the JUND-NECTIN4 axis (A and B) HT1376 and RT112 cells were transfected with the specified shRNA and plasmids for 72 h after puromycin screening. Then the cells were harvested for western blot analysis (A) and RT-qPCR analysis (B). (C) HT1376 and RT112 cells were transfected with the specified shRNA and plasmids for 72 h after puromycin screening. The cells were treated with continuous doses of EV for 24 h and harvested for CCK-8 assays to measure the IC50 values of EV. (D–I) HT1376 cells were transfected with the specified shRNA and plasmids for 72 h. After puromycin selection, the cells were collected for colony-formation assay (D), CCK-8 assay (E), western blot analysis (F), caspase-3 activity assay (G), and annexin V-FITC/PI assay (H and I). (J–L) HT1376 cells were transfected with designated shRNA for 72 h. After puromycin screening, these cells were injected subcutaneously into nude mice. These mice received or did not receive EV (intraperitoneal injection, 1 mg/kg once every 3 days five times in a row). Tumor image (J); tumor volume (K); tumor growth curve (L). Data were expressed as the mean ± SD and repeated five times. ns, not significant; *p < 0.05; **p < 0.01; ***p < 0.001. Data are representative of three independent experiments (B, D, E, G, and I). All data were expressed as the mean ± SD and repeated three times. ns, not significant; *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: For immunohistochemistry (IHC) of the tissuemicroarray (#U100Bl01), the following antibodies were used: YTHDC1 (#29441-1-AP, Proteintech, 1:1500 dilution), NECTIN4 (#67721-1-lg, Proteintech, 1:1500 dilution), and JUND (#AF6200, Affinity Biosciences, 1:1000 dilution).

Techniques: Transfection, shRNA, Western Blot, Quantitative RT-PCR, CCK-8 Assay, Selection, Colony Assay, Caspase-3 Activity Assay, Injection