Generation of stable cell lines expressing shRNA TEM4 constructs. (A) Immunofluorescence images of stable cell lines expressing shResistant TEM4^WT, TEM4^ΔABD, and TEM4^Y1216A. Scale bar = 5 μm for the main image and 1 μm for the zoomed image. (B and C) Relative actin intensity in TEM4-21 and TEM4-22 rescue cell lines. Between 35 and 90 cells were analyzed per condition from 3 replicate experiments. The bar represents the mean ± SD, and analysis of significance was performed using the Brown-Forsythe ANOVA test. (D and E) Circularity of TEM4-21 and TEM4-22 rescue cell lines. Between 35 and 90 cells were analyzed per condition from 3 replicates experiments. The bar represents the mean ± SD, and analysis of significance was performed using the Brown-Forsythe ANOVA test.