Figure 5
(A) Dasatinib treatment in H358 with or without TMPRSS4. Cells lacking TMPRSS4 are significantly more sensitive to dasatinib than controls. (B) Western blot analysis of DDR1 and TMPRSS4 in H358 cell clones. (C) Proliferation analysis by MTT in single and double KD in comparison with controls. (D) Morphological observation of double KD cells compared to controls. (E) Cell cycle analysis. The double KD clone showed reduced proportion of cells in the G2/M and S phases and arrest in the G0/G1 phase. (F) Western blot analysis showing cyclins and cell cycle-related proteins. Double KD cells lacked cyclin A, cyclin B1 and E2F1 protein expression. A remarkable increase in p21 was observed in these cells. (G) Tumor volume in mice injected with shDDR1 cells did not change with respect to controls. Upon administration of doxycycline, tumors lacking TMPRSS4 were significantly smaller than controls. Tumors from double KD cells underwent tumor regression. (H) Representative images of macroscopic and microPET images from the different groups (left panel). Quantification by microPET of maximum standardized uptake value (SUVmax) and metabolic tumor volume (MTV). In vitro experiments were repeated 3 times. Statistical analysis for SUVmax and MTV was not performed because most values in the double KD group were “0”. TMP: TMPRSS4.
