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. 2012 Aug;26(8):3230–3239. doi: 10.1096/fj.12-205609

Figure 1.

Figure 1.

Expression of E-cadherin (E-cad) in MLE-12 cells. A) Representative micrographs of MLE-12 cells stably expressing (sc-shRNA) and with reduced levels of SIK1 (SIK1-shRNA) (×100 view). B) Representative Western blot of SIK1 (1:1000), SIK2 (1:2000), SIK3 (1:2000), E-cadherin (1:400), LKB1 (1:500), GSK3β (1:1000), AMPK (1:500), and tubulin (1:1000). Equal amounts of protein (200 μg) were loaded in each lane. C) Expression of SIK1 and E-cadherin mRNA in cells lacking (SIK1-shRNA) vs. cells expressing SIK1 (sc-shRNA); results represent 3 experiments. D) Expression and localization of E-cadherin (1:100), β-catenin (1:100), and p120CAS catenin (1:200) were examined using fluorescent microscopy. Scale bars = 20 μm. E) p120CAS catenin mRNA expression levels in MLE-12 cells expressing- or lacking SIK1. Results represent 3 experiments. F) Right panel: expression of Snail1, Snail2, and TWIST. Left panel: expression of Zeb1 and Zeb2. Data are presented as percentage change or fold increase in cells lacking SIK1 compared wtih those expressing SIK1 (3 experiments). G) MLE-12 cells expressing or lacking SIK1 and cotransfected with the human variant of E-cadherin tagged with luciferase and either the human variant of SIK1 wild-type (SIK1-WT; open bars), the SIK1 T322A mutant (SIK1-T322A; solid bars), or the empty vector (pIRES). Luciferase was determined after 36 h expression. Values are expressed as percentage increase vs. cells expressing the empty vector. Results represents 4 experiments performed in triplicate. H) Expression of Snail2 (mRNA) in lung tissue from sik1+/+ and sik1−/− mice. Data are presented as arbitrary units (A.U.). Data represent 5 mice/group. I) Left panel: E-cadherin expression (mRNA and protein) in sik1+/+ and sik1−/− mice. Changes are expressed as percentage decrease in sik1+/+ vs. sik1−/− mice; n = 5 animals/group. Right panel: representative Western blot. Antibodies were used as in B. J) Left panel: expression of SIK2 (mRNA) in lung tissue from sik1+/+ and sik1−/− mice. Data are presented as arbitrary units; n = 5 animals/group. Right panel: expression of SIK2 (protein) in lung tissue from sik1+/+ and sik1−/− mice. Representative Western blot of 6 mice is shown. Bars represent means ± se.