Journal: Hypertension research : official journal of the Japanese Society of Hypertension

Article Title: Histone deacetyltransferase inhibitors Trichostatin A and Mocetinostat differentially regulate MMP9, IL-18 and RECK expression, and attenuate Angiotensin II-induced cardiac fibroblast migration and proliferation.

doi: 10.1038/hr.2016.54

Figure Lengend Snippet: Figure 1 Silencing HDAC1 and HDAC2 inhibits Ang-II-induced cardiac fibroblast proliferation and migration. (a) Expression of HDAC1 and HDAC2 at basal conditions was analyzed by immunoblotting. CF1, CF2: two independent cultures of primary cardiac fibroblasts. Cell extracts from human epithelial cells (HeLa) and mouse embryonic fibroblasts (NIH3T3) served as positive controls (n = 3). (b) Effect of Ang-II dose on activation of HDAC1 and HDAC2 in quiescent CF as analyzed by a colorimetric assay using nuclear protein extracts (100 μg) immunoprecipitated with HDAC1- or HDAC2-specific antibodies. Deacetylase activity was calculated in pmoles min −1 mg−1. Results are expressed as fold change from controls (n = 6). (c) Silencing HDAC1 or HDAC2 attenuates Ang-II-induced CF proliferation. CF transduced with lentiviral HDAC1 or HDAC2 shRNA (moi 0.5 for 24) were treated with Ang-II (10 −7 M) for 48 h, and proliferation analyzed by CyQUANT assay. (d) Silencing HDAC1 or HDAC2 inhibits Ang-II (10 −7 M for 12 h) -induced CF migration. Migration was analyzed using BioCoat Matrigel invasion chambers and MTT assay. (e) Knockdown of HDAC1 and HDAC2 was confirmed by immunoblotting (n = 3). Akt served as an off-target. (f) Silencing HDAC1 or HDAC2 failed to induce cell death. Forty-eight hours after lentiviral transduction, cell death was analyzed by an ELISA that quantifies mono and oligonucleosomal fragmented DNA in cytoplasmic extracts. (b, c, d, f) *Poat least 0.01 vs. untreated, †Poat least 0.05 vs. Ang-II (n = 6-12).

Article Snippet: Anti-vimentin (#sc-373717) antibodies used in fibroblast characterization, anti-caspase-3 (35 kDa, #sc-136219) antibodies and lentiviral GFP shRNA (#sc-45924-V) were from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).

Techniques: Migration, Expressing, Western Blot, Activation Assay, Colorimetric Assay, Immunoprecipitation, Histone Deacetylase Assay, Activity Assay, Transduction, shRNA, CyQUANT Assay, MTT Assay, Knockdown, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: RIPK1 activation mediates neuroinflammation and disease progression in multiple sclerosis

doi: 10.1016/j.celrep.2021.109112

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: RPL37A-VIC: Hs01102345 (TaqMan Gene Expression Assays) , ThermoFisher , N/A.

Techniques: Labeling, Recombinant, Sequencing, Selection, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Adjuvant, RNAscope, Gene Expression, Software