Partial rescue of perinuclear-concentrated networks in Krt14^−/− keratinocytes transfected with K14CF-C367. (A) Immunofluorescence-based imaging shows three modes of keratin filament network organization resulting from overexpression of GFP-tagged K14C367A and K14CF-C367 in Krt14^−/− mouse keratinocytes in primary culture. Bar, 10 µm. (B) Population frequency of each type of IF network as illustrated in A. Data are presented as mean ± SD from three independent experiments. (C) Still fluorescence images from video recordings of live mouse keratinocytes expressing GFP-K14CF-C367 (green channel). H2B-mCherry (red channel) was cotransfected to visualize nuclei. Seven time points from 0 to 6 h are shown (see Videos 4 and 5). Maximum intensity projections identify two different populations of mouse keratinocytes expressing the GFP-K14CF-C367: with (w/periN) or without (w/o periN) perinuclear-concentrated networks (45% vs. 55%, respectively). Bar, 10 µm. (D) 2.5-dimensional views of z-stacked images corresponding to the two-dimensional images at time 0, 3, and 6 h in C.