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. 2019 Feb 7;9(5):1247–1263. doi: 10.7150/thno.29884

Figure 3.

Figure 3

Binding and internalization of PSP and PSPS in PSMA+ or PSMA- CRPC cells. Specific binding of PSP with PSMA+ prostate cancer cell lines analyzed by (A) FCM assays and (B) IF assays (400x). PSMA+ prostate cancer cell lines C4-2 (FCM and IF), PC-3-PSMA+ (FC), LNCaP (FCM) and PSMA- prostate cancer cell lines PC-3 (FCM) and DU145 (FCM)were incubated with 50nM PSP, PSMAb (positive control) or human IgG (negative control) followed by PE-IgG-Fc antibody. Specific internalization of PSP into PSMA+ CRPC cells analyzed by (C) FCM and (D) IF assays (x 400). For the FCM assay, C4-2 or PC-3 cells were incubated with 100nM PE-PSP and PE-PSMAb for 1 h at 4°C or 37°C followed by trypsin digestion. For IF assay, C4-2 cells were incubated with 100nM PSP, PSMAb (positive control), or human IgG (negative control) for 3 h at 37°C followed by PE-conjugated anti-human IgG Fc antibody. Internalization of FAM-siRNA into PSMA+ CRPC cells analyzed by (E) FCM assay and (F) IF assay (x 400). C4-2 (FCM and IF) or PC-3 (FCM) cells were incubated with 100 nM PSPS. DAPI was used to visualize the nuclei. Scale bars = 20 μm. Representative results of 3 independent experiments are shown.

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