Fig. 6: TmpC-CKAP4 stabilized HIF-1α by binding RanBP2 and induced autophagy by binding NBR1.

a Immunoprecipitation of CKAP4 followed by HPLC-MS identified CKAP4-bound proteins. b Co-IP and immunoblotting detection showed the interaction between RanBP2 and CKAP4. c Co-IP and immunoblotting detection showed the interaction between HIF-1α and SUMO1. d TAK981 decreased the expression of HIF-1α and HK2. The samples derive from the same experiment but different gels for HIF-1α and ACTB, another for HK2 were processed in parallel. e siRanBP2 decreased the expression of HIF-1α. f Co-IP and immunoblotting detection showed the interaction between NBR1 and CKAP4. g Co-IP and immunoblotting detection showed the binding of P62 with CKAP4 and NBR1. h siNBR1 increased P62 and decreased LC3B II. i siNBR1 decreased the P62 bodies (n = 5 cells per group). The samples derive from the same experiment but different gels for NBR1 and ACTB, another for P62 and LC3B were processed in parallel. j–l Representative immunofluorescence images (j) and quantification (k, l, n = 15 cells per group) showing siCKAP4 inhibited the co-localization of NBR1 with P62 bodies. The ratio quantified by comparing the number of NBR1-positive P62 bodies to all P62 bodies per cell. Scale bar: 10 μm. m A schematic diagram illustrating the mechanisms of P. micra TmpC promoted OSCC metastasis, created in figdraw.com. Data were shown as mean ± SD. Blots are representative of at least two biological replicate experiments with similar results (b–h). One-way ANOVA with Turkey’s test was used to examine the statistical significance between groups.