α histone h3

The mouse mAb S16 was produced by Sugawara et al.^{27 (link)}. BALB/c mice were immunized with 50 μg purified HEF proteins of C/Ann Arbor/1/50 in Freund’s complete adjuvant thrice at weekly intervals. The hybridoma cells secreting antibodies to HEF were screened using enzyme-linked immunosorbent assay (ELISA). The isotype of S16 was classified as IgG2a using double immunodiffusion.
For western blotting, α-mouse ACAA2 (ab128911; Abcam, Cambridge, UK), α-GAPDH (#5174; Cell Signaling Technology, Danvers, MA, USA), α-HSP60 (#12165; Cell Signaling Technology), α-histone H3 (#4499; Cell Signaling Technology), α-Flag M2 (F1804; Sigma-Aldrich, St. Louis, MO, USA), and α-mouse albumin (#4929; Cell Signaling Technology) polyclonal antibodies (pAbs) were used as the primary antibodies. Horseradish peroxidase (HRP)-conjugated secondary antibodies against mouse and rabbit IgG were purchased from Bio-Rad (Hercules, CA, USA). The pAb against mouse ACAA2 (TA321746; OriGene, Rockville, MD, USA) and HRP-conjugated goat α-mouse IgG2a antibody (Southern Biotech, Birmingham, AL, USA) were used for ELISA. The IgG2a isotype control (BioLegend, San Diego, CA, USA) was used as a control antibody for the indirect immunofluorescence assay and injection of S16 into mice.

Plasmids were generated and sequence-verified following standard cloning procedures. GCaMP6f was described in ref. ^{20 (link)}. AC-GFP and Lamin-Chromobody-GFP were from ChromoTek. nAC-mCherry and Lamin-Chromobody-mCherry were constructed by exchanging the GFP with mCherry. nAC-GFP and nAC-mCherry were ligated into pWPXL for lentiviral transduction. INF2-CAAX full length in pEGFP-C1 was from Henry N. Higgs. BFP-INF2-CAAX was cloned by exchanging the EGFP in pEGFP-C1 vector with TagBFP2. Calmodulin and its mutant was a gift from Tim Plant. mCherry-CaMBP4 and the control plasmids were published by Monaco et al.^{24 (link)}. Cytoplasmic AC-mCherry-NES was cloned by adding an NES (nuclear export signal) after mCherry. INF2-DID (aa 32–266) and INF2-DAD (aa 965–1249) were cloned into EFpLink vector. α-Gα₁₂ and α-Gαq were from Santa Cruz. α-INF2 was from Proteintech. α-Tubulin and α-histone H3 were from Cell Signaling Technology. α-Flag conjugated with HRP and α-myc conjugated with HRP were from Sigma. α-GFP (B2) was from Santa Cruz.

The whole cell proteins of MDA-MB-231/MDA-MB-468 cells were extracted from cells using RIPA buffer (Beyotime, Shanghai, China) and the nuclear extracts were prepared with a NE-PER Nuclear and Cytoplasmic Extraction Kit (Pierce Biotechnology, Rockford, USA). Western blots were probed with α-phospho LRP6 (Ser1490), α-LRP6, α-β-catenin, α-E-cadherin, α-N-cadherin, α-Vimentin, α-Snail, α-Histone H3, α-c-Myc, α-cyclin D1, α-Axin, α-CD44, α-HIF-1a, α-Glut1 (Cell Signaling Technology, USA), α-VEGFA (Abcam, Cambridge, UK), α-β-actin.