Journal: Advanced Science

Article Title: Targeting NSUN2‐Mediated m 5 C Modification Attenuates Chondrocyte Senescence and NLRP3 Activation in Osteoarthritis

doi: 10.1002/advs.76370

Figure Lengend Snippet: Ca 2+ promotes the upregulation of NSUN2 expression by targeting SP1, forming a positive feedback loop. (A, B) Relative protein expression level and quantification of NSUN2 in Primary human chondrocytes (HPCs) treated with different concentrations (0, 50, 100, and 200 µM for 24 h) or different durations (100 µM for 0, 1, 12, and 24 h) of histamine. (C) Relative mRNA levels of NSUN2 in HPCs treated with different concentrations (0, 50, 100, and 200 µ m for 24 h) of histamine. (D) Relative protein expression level and quantification of IP3R3 and NSUN2 from HPCs treated with histamine following in the presence of DMSO or 2‐APB (50 µM). (E) Relative m 5 C level and quantification in HPCs treated with Histamine following in the presence of DMSO or 2‐APB (50 µM). (F) Eight candidate transcription factors (TFs) were identified through integrated analysis of three prediction databases: JASPAR, hTFtarget, and KnockTF. (G, H) Relative mRNA and protein levels of NSUN2 in HPCs treated with histamine alone or combined with Plicamycin (25 nM) or 10058‐F4 (50 µM) (Data was presented as mean ± SD, *** p < 0.001, ** p < 0.01, * p < 0.05, ns, no significance).

Article Snippet: 2‐APB (524‐95‐8), BAPTA/AM (126150‐97‐8), Plicamycin (18378‐89‐7), 10058‐F4 (403811‐55‐2) were purchased from MCE; NSUN2 (1:10000, 20854‐1‐AP), p21 (28248‐1‐AP, 1:1,000 dilution), TNF‐α (17590‐1‐AP, 1:1000), NLRP3 (68102‐1‐Ig, 1:2,000 dilution), ASC (10500‐1‐AP, 1:5,000 dilution), Caspase1 (22915‐1‐AP, 1:2,000 dilution), IL‐1β (16806‐1‐AP, 1:2,000 dilution), IP3R3 (20729‐1‐AP, 1:1,000 dilution), YBX1 (20339‐1‐AP), GAPDH (HRP‐60004, 1:5,000 dilution) antibodies were purchased from Proteintech (Wuhan, China), COL2A1(GB11021‐100, 1:1000), NEK7 ( GB112643 ‐100, 1:1,000), p16 ( GB111605 ‐100, 1:1,000 dilution) and MMP13 (GB11247‐1‐100, 1:1,000 dilution) antibodies were purchased from Servicebio (Wuhan, China).

Techniques: Expressing

Journal: Molecular Oncology

Article Title: Inhibition of CDK 9 enhances AML cell death induced by combined venetoclax and azacitidine

doi: 10.1002/1878-0261.70124

Figure Lengend Snippet: mTOR, c‐MYC, and MCL‐1 contribute to AraC‐resistant AML cell survival. (A) gsea comparing proteomics results of MV4‐11 and MV4‐11/AraC‐R cells (published in reference ) to the Hallmark gene set. (B) Whole cell lysates from AML cell lines, J000106565 PDX cells, and KCI48595 primary blasts were subjected to western blot and probed with anti‐p‐S6 and anti‐β‐actin antibodies. Densitometry measurements normalized to β‐actin and then compared to parental cell lines are shown below the corresponding blot. (C–H) AraC‐resistant AML cell lines were treated with 10058‐F4 or AZD5991 for 24 h; parental and AraC‐resistant AML cell lines were treated with rapamycin for 24 h. c‐MYC, MCL‐1, or p‐S6 protein levels, from whole cell lysates, were analyzed by western blot (panels C, E, and G). Densitometry measurements normalized to β‐actin and then compared to the control are shown below the corresponding blot. Annexin V‐fluorescein isothiocyanate (FITC)/propidium iodide (PI)‐stained cells were analyzed by flow cytometry (panels D, F, and H). Results are graphed as mean ± SEM from one experiment in triplicate. *** P < 0.001 compared to the control. The unpaired t ‐test was used to determine statistical significance. (I) AraC‐resistant AML cell lines were treated with AZD5991, 10058‐F4, or rapamycin, alone or in combination, for 24 h, then stained with annexin V‐FITC/PI and analyzed by flow cytometry. Results are graphed as mean ± SEM. *** P < 0.001 compared to the control. ### P < 0.001 compared to all corresponding single‐drug treatments. $$$ P < 0.001 compared to all corresponding 2‐drug combinations. One‐way ANOVA followed by Bonferroni's post hoc test was used to determine statistical significance. Western blotting and annexin V‐FITC/PI staining and flow cytometry analysis experiments were repeated in biological replicates.

Article Snippet: AZD4573, 10058‐F4, IACS‐010759, and AZA were purchased from Selleck Chemicals (Houston, TX, USA).

Techniques: Western Blot, Control, Staining, Flow Cytometry

Journal: Cell stem cell

Article Title: Deterministic Somatic Cell Reprogramming Involves Continuous Transcriptional Changes Governed by Myc and Epigenetic-Driven Modules

doi: 10.1016/j.stem.2018.11.014

Figure Lengend Snippet: Key Resources Table

Article Snippet: c-Myc inhibitor 10058-F4 , Axon Medchem , #2222.

Techniques: Recombinant, Protease Inhibitor, Sample Prep, Methylation, Transgenic Assay, Negative Control, Software