Extended Data Figure 1: Proliferation of cone-like cells after Rb depletion in dissociated FW19 retina.

a, Decreased Rb protein in L/M-opsin⁺ or TRβ2⁺ cells (arrows) on days 5 or 23, and decreased RB1 RNA or Rb protein on day 4 after shRB1-733 transduction. b, Cone arrestin⁺, CRX⁺ cells (arrows) with or without Ki67 co-expression. c, Ki67⁺ and cone arrestin⁺ cells first detected 9 or 14 days after transduction in two experiments. d–f, Co-staining of Ki67 with RXRγ and CRX at 14 days (d), with cone arrestin and CRX at 14 days (e), or with L/M-opsin and CRX at 23 days (f) after transduction with shRB1-733 or a scrambled control. g, Percentage of cells co-expressing Ki67 with L/M-opsin and CRX, RXRγ and CRX, or cone arrestin and CRX, 23 days after transduction. h, Prevalence of cells co-staining for L/M-opsin and CRX, RXRγ and CRX, or cone arrestin and CRX, 23 days after transduction. i, Ki67 not detected in cells expressing markers of rods (NRL), ganglion cells (BRN-3), bipolar cells (strong CHX10), or horizontal cells (PROX1) 14 days after transduction. j, Co-expression of Ki67 with markers of RPCs (nestin, white arrows) or Müller glia (CRALBP or SOX2), but not in PAX6⁺, nestin⁻ ganglion, amacrine or horizontal cells (yellow arrows) 14 days after transduction. k, l, EdU incorporation in cells expressing markers of cones (cone arrestin and CRX or RXRγ and CRX, yellow arrows in l) but not in cells expressing markers of rods (CNGA1, CNGB1), bipolar cells (CHX10, CRX), or ganglion, horizontal or amacrine cells (syntaxin) (white arrows in l) 14 days after transduction. Black lines above labels demarcate distinct fields. m, Co-staining of phosphohistone H3 (PH3) with cone arrestin and CRX 23 days after transduction. n, Apoptosis marker CC3 in cells expressing RPC and glial marker nestin 14 days after transduction with RB1-directed shRNAs (yellow arrow) but not with scrambled control (white arrow). Values and error bars are mean and s.d. of triplicate assays. Scale bars, 20 μm. Data are representative of at least two independent experiments.