Fig. 4

FLJ enhances CRPC cell proliferation by inducing autophagy in response to trace levels of androgen. A Immunofluorescence was used to detect LC3B expression, an autophagy marker, following FLJ knockdown in cells, with a scale bar of 25 μm. B The mean fluorescence intensity of LC3B in Figure A was quantified. C The response of FLJ knockdown to trace androgen stimulation was evaluated using the CCK8 assay. D Western blot analysis was conducted to assess the regulation of autophagy markers after FLJ knockdown under conditions of androgen deprivation or the addition of trace androgens. E Transmission electron microscopy (TEM) was used to observe autophagosome regulation after FLJ knockdown under androgen deprivation or the addition of trace androgens, with a scale bar of 500 nm. F Quantification of autophagosomes in Fig. 4D was performed. G The effects of FLJ knockdown on cell proliferation and viability were assessed using the CCK8 assay, following autophagy induction by trace androgens, under normal culture conditions (left) and in vitro castration-simulated conditions (right). H, I The colony-forming ability after FLJ knockdown was evaluated using a colony formation assay, following autophagy induction by trace androgens, under normal culture conditions (top) and in vitro castration-simulated conditions (bottom). J, K The IC50 value of ENZA was determined using the IC50 assay after FLJ knockdown and autophagy induction by trace androgens. L The effect of 20 μM ENZA on cell viability over 0–96 h was evaluated using the CCK8 assay after autophagy induction by trace androgens. M The impact of FLJ knockdown on cell viability was assessed using the CCK8 assay at 48 h after treatment with 20 μM and 50 μM ENZA, following autophagy induction by trace androgens