mouse anti phh3  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: Targeted H3K9 acetylation at lncSox1 promoter by cell type-specific epigenome editing promotes intermediate progenitor proliferation in developing mouse cortex

doi: 10.1101/2024.11.05.621758

Figure Lengend Snippet: (A) Micrographs showing immunostaining analysis of pHH3, TBR2, GFP in the E15.5 mouse cortex electroporated at E13.5 with sgLncSox1 and sgControl plasmids. Inserts are high magnifications of cells stained with the indicated markers and display M-phase status. Yellow arrows point to TBR2+ cells with mitotic figures and white arrows point to those without mitotic figures. (B) Statistical quantification of proliferating TBR2-expressing cells in M-phase in the sgLncSox1- and sgControl-treated E15.5 cortex. (C) Micrographs showing immunostaining analysis of BrdU, TBR2, GFP in the E15.5 mouse cortex electroporated with sgLncSox1 and sgControl plasmids. Inserts are high magnifications of cells stained with the indicated markers and display S-phase status. Yellow arrows point to TBR2+ cells with active DNA replication and white arrows point to those without active DNA replication. (D) Statistical quantification of proliferating TBR2-expressing cells in S-phase in the sgLncSox1- and sgControl-treated E15.5 cortex. (E) Micrographs at low (upper panels) and high (lower panels) magnifications showing GFP, mCherry, and NeuN immunostaining in the E18.5 mouse cortex electroporated at E13.5 with sgLncSox1 and sgControl plasmids. Red rectangle demarcates zoomed area of the cortical plate shown in the lower panel micrographs for quantitative analysis. White arrows point to mCherry/NeuN cells. Their lack of GFP expression indicate they are derived from non-TBR2-expressing cells or direct neurogenesis. Cells with GFP/mCherry/NeuN staining are derived from TBR2+ cells via indirect neurogenesis. (F) Statistical quantification of the proportion of cells undergoing direct or indirect neurogenesis in the sgLncSox1- and sgControl-treated E18.5 cortex. Values are presented as mean ± SEM (** p < 0.01). Scale bars = 50 μm. (G) Schema illustrating how lncSox1 expression in the normal mouse cortex is augmented by its promoter region H3K9ac enrichment leading to intermediate progenitor pool expansion, which has implication for expansion and folding of the epigenetically manipulated mouse cortex. The outcome of such epigenetic manipulation phenocopies the human cortical with possibly evolutionary basis. Abbreviations: Ac, H3K9ac mark; IPCs, intermediate progenitor; VZ, ventricular zone; SVZ, subventricular zone; IZ, intermediate zone; CP, cortical plate.

Article Snippet: Monoclonal (mAb) and polyclonal (pAb) primary antibodies were commercially sourced and listed as follows: GFP chick pAb (1:400; Abcam), mCherry/RFP pAb (1:1000, biomol/Rockland), mCherry/RFP mAb (1:1000, biomol/Rockland), NeuN mouse mAb (1:200, Chemicon), HuCD mouse mAb (1:20; Invitrogen), Pax6 rabbit pAb (1:200; Covance), Pax6 mouse mAb (1:100; Developmental Studies Hybridoma Bank), TBR2 rat 923 mAb (1:200; eBioscience), TBR2 rabbit pAb (1:200; Abcam), H3K9ac pAb (ChIP-seq, Millipore), pHH3 mAb (1:50; Cell Signaling), BrdU rat pAb (1:100; Abcam), BrdU mouse mAb (1:40; CalTag), Tuj mAb (1:200; Chemicon).

Techniques: Immunostaining, Staining, Expressing, Derivative Assay