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. 2022 Sep 15;9:1006497. doi: 10.3389/fmolb.2022.1006497

FIGURE 4.

FIGURE 4

Effects of paclitaxel on cell proliferation, cell death and involved proteins in mature brain organoids. (A,B) Immunohistochemistry analyses of cell proliferation in brain organoids treated with paclitaxel (EC50: 153 nM and EC90: 1,084 nM) for 14 h with a subsequent culture in fresh media for 15 h and 3 h in BrdU containing medium (32 h sampling time point) or with medium (MC) or vehicle (VC) as controls (A), demonstrated a 74% or 86% lower number of BrdU + cells in organoids treatedwith EC50 or EC90 respectively compared to the VC (n = 3) (B). (C,D) Analysis of apoptosis via TUNEL assay in sections of the same brain organoids as used for proliferation assessment (C) showed a 2.9-fold and 10.9-fold increase in apoptotic cells in brain organoids treated with EC50 or EC90 paclitaxel respectively compared to the VC (n = 3) (D). (E,F) Western blot analysis of organoids treated with paclitaxel for 14 h (EC50: 153 nM and EC90: 1,084 nM) with subsequent culture in fresh media for 10 h or 18 h (24 and 32 h sampling time points) show lower neuronal calcium sensor protein 1 (NCS-1) (n = 2) (E) and higher cleaved caspase 3 (n = 3) (F) abundance compared to the VC in a time- and dose-dependent manner. Histologic data are normalized to VC, Western blot data are normalized to a normalization sample. All data are shown in means and standard deviation.