a, Astrocytes (green) were transfected with shRNAs against rat NL1 (shrtNL1), mouse/rat NL2 (shNL2), and/or rat NL3 (shrtNL3) ± HA-tagged, shRNA-resistant NL plasmids (red) and co-cultured with neurons (not visible). b-e, Quantification of astrocyte complexity for conditions in (a). f, Image of an astrocyte transfected with shrtNL1 (green) and HA-tagged NL1-SWAP (red) in co-culture with neurons (not visible). g, Quantification of astrocyte complexity for (f). h, Images of shCtrl-transfected astrocytes (red) in co-culture with shNrx1/2 or shScr lentivirus-transduced neurons (green). i, Quantification of astrocyte complexity for (h). b-e, i Data represented are from individual experiments with 3 biological replicates. Similar results were obtained in 3 independent experiments. n>20-25 cells/condition/experiment. j, Nanofibers were coated with Fc-tagged proteins to model a neuronal scaffold. Images of EGFP-transfected astrocytes (green) cultured on Fc-protein-coated nanofibers (red). k, Quantification of astrocyte complexity for (j). Data represent 1 experiment with 4 biological replicates. n>25 cells/condition. ANCOVA (b-e, g, i, k). Data are means ± s.e.m. Scale bar, 10 μm.