Dapi staining reagent

The cover glass was placed in the 6‐well plate. Intestinal macrophages were inoculated and stimulated with LPS for 12 h, fixed with 4% paraformaldehyde (PFA) for 10 min, washed with PBS for three times and permeabilized with 0.2% Triton X‐100 for 10 min, blocked with 2% BSA for 30 min, incubated with MD2 monoclonal antibody (Abcam dilution 1:300) for 1 at room temperature, incubated with IgG antibody (Abcam) for labelling, added with 0.5 μg/ml DAPI staining reagent (Solarbio) for nuclear staining, washed with PBS for two times, mounted and observed under fluorescence microscope.

UMUC‐3 and 5637 cells were respectively plated onto 12 wells at a density of 5.0 × 10⁴ cells per well at 37°C in a 5% CO₂ incubator. After 24 h, treated with packaged lentiviruses paCas9‐Survivin and paProtacL‐Survivin separately or in combination for 48 h, and then illuminated by 1.2 W/m² blue light for another 48 h. Immediately after, the medium was aspirated and rinsed gently twice with pre‐chilled PBS. Subsequently, the living and dead cells were identified by calcein/PI cell viability/cytotoxicity Assay Kit (Beyotime) and DAPI staining reagent (Solarbio), consistent with the manufacturer's guidelines. Then, the cells were examined by a fluorescent microscope (Olympus). FITC channel (λex 488 nm and λem 525 nm), PI channel (λex 535 nm and λem 615 nm) and DAPI channel (λex 364 nm and λem 454 nm).