Fig 2: Lacripep re-establishes functional corneal nerve-epithelial interactions during dry eye disease progression(A) Schematic of the treatment regimen showing data collection time points for RNA-seq and tissue analyses.(B–C) Immunofluorescent analysis (B) and quantification (C) of regenerating nerves in the corneal epithelium at day 7. The graph in (C) shows the relative proportion of newly regenerating GAP43+ nerves in the central cornea. Scale bars, 50 µm.(D) 3D reconstruction of whole-mount immunofluorescent images of newly regenerating (GAP43+) and existing (TUBB3+) intraepithelial nerve terminals at days 7 (upper panel) and 15 (lower panel) in the corneal epithelium. Scale bars, 25 µm (IF), 10 µm (3D reconstruction).(E) Graph showing relative proportion of GAP43+, GAP43 + TUBB3+, and TUBB3+ nerve terminals in the central cornea at day 7. NUC, nuclei. All data are expressed as mean + SD. n > 4 mice per group.

Fig 3: EXSISERS can be programmed with different reporters and effectors.a,b, HaloTag was flanked by intein-CC domains and type I and type II transmembrane domains (TMDs), resulting in a surface functionalization with a HaloTag inserted into exon 10 of MAPT. c, Cells were incubated with chloroalkane-functionalized Alexa Fluor 660 (3.5 µM) for 10 min at 37 °C and washed with HBSS. d, Anti-pan-tau immunoblot showed the typical tau pattern indicating scarless excision. Data shown represent one independent experiment. e, Epifluorescence microscopy image of anti-pan-tau immunocytochemistry of EXSISERSMAPT:10Halo cells after MAPT induction showing filamentous tau staining. Data shown represent 2 independent experiments f, AF660 live-cell-staining after MAPT-induction resulted in a distinct membrane staining. Data shown represent 2 independent experiments. g, scAvidin was used as a high-affinity binding domain for biotinylated compounds. Test constructs for both designs contained mNG-NLS as surrogate exteins, which are expected to be spatially decoupled after the excision of the intein-domains. h,i, Surface localization of scAvidin or HaloTag in transfected HEK293T was confirmed by live-staining with biocytin-AF594 and chloroalkane-AF660, while mNeonGreen-NLS (mNG-NLS) was detected in the nucleus. Data shown represent one experiment from 2 cell culture wells. j, The binding moiety of the membrane anchors was exchanged for NLuc flanked by furin sites (FS), such that the Golgi-resident furin can cleave off NLuc and releases it into the supernatant. k, NLuc activity was measured over time in the supernatant of cells expressing the sheddable NLuc without/with furin sites. Data shown represent the mean values ± s.d. (n = 3 biological replicates). *** denotes p < 0.001 of selected Bonferroni MCT after two-way ANOVA with repeated measures (full statistical results in Supplementary Table 1). l, Intein-flanked mNG was inserted into the mouse Tubb3 gene. m, Immunoblot confirms successful intein splicing of mNG. Data shown represent 7 independent experiments. n, Mouse N2a cell line with insertion of the intein-mNG reporter into Tubb3 shows typical Tubb3 filaments via immunocytochemistry. The fluorescent mNG signal (inset) was observed throughout the cell, indicating successful post-translational splicing. Data shown represent 2 independent experiments. Unprocessed blots and numerical source data are provided in Source Data Extended Data Fig. 8.Source data