inca 6  (MedChemExpress)

Journal: Scientific Reports

Article Title: Calcineurin/NFATc1 pathway represses cellular cytotoxicity by modulating histone H3 expression

doi: 10.1038/s41598-024-65769-9

Figure Lengend Snippet: NFATc1 downregulates the expression of histone H3 ( A ) Isolation of factors that regulate histone H3 mRNA expression. From a comprehensive database analysis, we isolated three groups of indicators: Transcription factor that binds to the transcription start site (TSS) ± 500 bp of histone H3, factors known as transcription factors, factors interact with calcineurin from the BioGRID database. ( B ) MCF7 cells were treated with DMSO or 50 μM FK506 for 24 h and fractionated into cytosol and nuclear fractions. Each fraction was subjected to immunoblotting using the indicated antibodies. ( C ) Lentivirus-infected MCF7 cells were cultured in the presence of Dox to induce the expression of NFATc1 and NFATc3 shRNA. Cell lysates were prepared and analyzed by immunoblotting with the indicated antibodies. D) RT-qPCR analysis of H3.1, H3C10, and H3.3 mRNA in MCF7 cells expressing NFATc1, NFATc3, and control shRNAs. For qPCR analysis, TBP was used as a control for the normalization of mRNA data. Data are expressed as mean ± SEM of three independent experiments. ns: not significant, ****P < 0.0001, ***P < 0.001(one-way ANOVA) E,F,G) RT-qPCR analysis of H3.1, H3.3, and CENPA mRNA in MCF7 cells treated with the indicated concentrations of INCA-6 for 27 h. For qPCR analysis, 18S rRNA was used as a control for the normalization of mRNA data. Data were analyzed as described in D. ns: not significant, ***P < 0.001,**P < 0.01 (one-way ANOVA) Full uncropped blots are available in Supplemental Figure S5.

Article Snippet: The cells were treated with FK506 (063–06,191; Wako), CN585 (207,003; Merck), INCA-6 (ab145864; abcam), ionomycin (095–05,831; Wako), and verapamil (222-00,781; Wako).

Techniques: Expressing, Isolation, Western Blot, Infection, Cell Culture, shRNA, Quantitative RT-PCR, Control