Alexa fluor 488 conjugated donkey anti goat igg h l

Cells were fixed with 4% PFA in PBS for ~5 min at room temperature. After blocking with 20% AquaBlock (#PP82; East Coast Bio, North Berwick, ME, USA, https://eastcoastbio.com/) for 30 min at room temperature, cells were incubated overnight at 4°C with goat anti-Pdx1 antibody (1:100; #ab47383; Abcam, Tokyo, Japan, http://www.abcam.com/), goat anti-insulin antibody (1:100; #ab7842; Abcam), or rabbit anti-C-peptide antibody (1:100; #4593; Cell Signaling, Tokyo, Japan, http://www.cellsignal.com/). This was followed by incubation for 1 h at room temperature with Alexa Fluor 488- conjugated donkey anti-goat IgG H&L (1:200; #ab150129; Abcam), FITC-conjugated anti-goat IgG (1:250; #ab6904; Abcam), or Alexa Fluor 647-conjugated anti-rabbit IgG (1:250; #4414; Cell Signaling), respectively. A medium containing DAPI (#H-1200; Vector Laboratories, Burlingame, CA, USA, https://www.vectorlabs.com/default.aspx) was used for mounting and nuclear staining.

The following primary antibodies were used in this study: HDAC1 (histone deacetylas 1) mouse monoclonal antibody (mAb) (cat#5356, 1:1000), HDAC2 mouse mAb (cat#5113, 1:1000), HDAC3 mouse mAb (cat #3949, 1:1000), HDAC4 rabbit mAb (cat #7628, 1:1000), Histone H3 rabbit mAb (cat#4499, 1:1000), Acetyl-Histone H3 (Lys18) (H3K18ac) rabbit mAb (cat#13998, 1:1000), β-Actin rabbit mAb (cat#4970, 1:1000), HRP (horseradish peroxidase), labeled anti-mouse IgG (cat#7076, 1:3000) and HRP labeled anti-rabbit IgG (cat#7074, 1:3000), were all purchased from Cell Signaling Technology (Beverly, MA, USA). β-Tubulin rabbit polyclonal antibody (cat# AC015) was purchased from Abclonal Science Inc (Woburn, MA, USA). The HAT inhibitor inhibitors, including Anacardic acid (AA) [41A7236], as well as HDAC inhibitor trichostatin A (TSA) (cat#8552), were ordered from Sigma-Aldrich (St. Louis, MO, USA). Goat anti-BoHV-1 serum (cat# PAB-IBR) was purchased from VMRD Inc (Pallman, WA, USA). Alexa Fluor 488^®-conjugated donkey anti-goat IgG H&L (ca# ab150129) was purchased from Abcam (Cambridge, England).