Fig. 2

Starvation-Induced Autophagy Promoted the Progression of BLCA. a After normal or HBSS treatment, T24 cells were fixed, and the formation of autophagosomes was observed by transmission electron microscopy. Autophagosomes are marked by red arrows (scale bar, 2 mm). b Immunofluorescence staining of LC3B was performed in T24 cells with normal or HBSS treatment, and the aggregation of LC3B represented the formation of autophagosomes. c Western blotting was used to detect the expression of LC3B-I/II and SQSTM1/P62 to verify the efficiency of autophagic flux inhibition by CQ in different strains of cells and different treatments. d CCK-8 assay demonstrated the viabilities of T24 cells after treatment, including normal, normal supplemented with CQ, starvation only and starvation supplemented with CQ (n = 3, Mann–Whitney test, P* < 0.05, P** < 0.01, P*** < 0.001). e Images of tumours in nude mice bearing T24 cells treated with normal, normal supplemented with CQ, starvation only and starvation supplemented with CQ. f Tumour volume was measured after different treatments in nude mice every 3 days. The arrow indicates the timing of intraperitoneal administration. g and h Cell Transwell migration and invasion assays were performed in T24 and UM-UC-3 cells with different treatments (n = 3, unpaired Student’s t test, P* < 0.05, P** < 0.01, P*** < 0.001)