FIGURE 1.

Flow cytometry screen identifies novel markers for neurotoxic TIC-induced reactive astrocytes. (A) Representative images of major timepoints in hiPSC differentiation protocol into astrocytes. Between days 66 and 75 astrocytes are purified using CD49f (representative flow cytometry contour plot shown). Immunofluorescence staining for GFAP in unstimulated and TIC-induced reactive astrocytes highlights the change in morphology caused by proinflammatory stimulation. (B) Representative images of GFAP immunofluorescence staining in unstimulated and TIC-induced reactive astrocytes at high magnification. We used inverted grayscale to emphasize the changes in morphology caused by proinflammatory stimulation. (C) Schematic of flow cytometry screening of 361 PE-conjugated antibodies identifies six novel markers of TIC-induced reactive astrocytes, listed in the table. (D) Flow cytometry detection of each marker from the screen is shown as histogram plot comparing TIC-induced reactive (red) to unstimulated (gray) astrocytes.