Overexpression of ZNF416 drives lung fibroblast proliferative, contractile, and matrix synthetic activation.
(a) qPCR of primary human lung fibroblasts stably overexpressing ZNF416 (O.E. ZNF416) or an empty vector control. n = 3 independent biological replicates. (b) Violin plot of overexpressing ZNF416 primary human lung fibroblasts and empty vector controls in the presence or absence of TGFβ. Solid lines represent median. Dotted lines represent upper and lower quartiles. (c) Ki67 nuclear intensity, assessed by quantitative immunofluorescence, in primary human lung fibroblasts following ZNF416 overexpression. n = 4 biological replicates. Fold change in cell counts of primary human lung fibroblasts between 1 and 3 d following overexpressing ZNF416 or empty vector control. Data normalized to the cell counts at day 1 to observe proliferation. n = 3 biological replicates. (d) Phalloidin and DAPI staining with quantification of fibroblasts overexpressing ZNF416 and empty vector. Scale bars represent 50 µm. (e) Deposition of collagen I and fibronectin by primary human lung fibroblasts overexpressing ZNF416 or empty vector control following 3 d of culture. Data normalized to cell counts per image. *, P < 0.05; **, P < 0.01; ****, P < 0.0001, evaluated by unpaired t test (a, c, and d) or one-way ANOVA (b). All error bars represent SEM. E.V., empty vector control; F.C. Rel., fold change relative; O.E., overexpressing.