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. 2019 Nov 18;130(1):157–170. doi: 10.1172/JCI127144

Figure 6. EMP2 regulates adhesion molecules and TEM through caveolin-2.

Figure 6

(A) Caveolin 1 (Cav1) and Cav2 mRNA was quantified by RT-qPCR in lungs of naive Emp2+/+ and Emp2–/– mice (n = 3/genotype). (B) Cav1 and Cav2 mRNA was quantified by RT-qPCR in Calu-3 cells after stable EMP2 silencing with 2 lentiviral shRNA constructs, or treatment with scrambled (scr) lentiviral shRNA control (4 biological replicates). (C) EMP2 was ectopically expressed in CMT 64 cells; empty vector served as a control. Emp2, Cav1, and Cav2 were then quantified by RT-qPCR (4 biological replicates). (D) Stable EMP2 knockdown (KD) or scrambled control (WT) Calu-3 cells underwent lentiviral shRNA silencing of Cav-2, or parallel treatment with scr vector. Cav2 mRNA was quantified by RT-qPCR (left). Cell surface CD47, ICAM-1, and β3-integrin were quantified by flow cytometry and are depicted as relative MFI (right). (E) Stable EMP2 KD or WT Calu-3 cells were transduced with doxycycline-inducible (dox-inducible) Cav-2 shRNA constructs and then left untreated or treated with doxycycline. Cav-2 silencing was confirmed by RT-qPCR (left). Human neutrophil (PMN) transmigration to fMLP across Cav-2–silenced and unsilenced monolayers was quantified (right; n = 6–8 wells per condition, normalized across 2 independent experiments). Data are mean ± SEM. A–D are representative of at least 3 independent experiments. *P < 0.05; **P < 0.01 analyzed using unpaired 2-tailed Student’s t test in A and C–E, or 1-way ANOVA followed by Dunnett’s test in B.