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. Author manuscript; available in PMC: 2018 Oct 5.
Published in final edited form as: Cell Stem Cell. 2017 Sep 28;21(4):472–488.e10. doi: 10.1016/j.stem.2017.08.014

Figure 4. Putative iAEC2 Lamellar Bodies Function to Synthesize and Secrete Surfactant.

Figure 4

(A) Cellular compartments in which proSFTPB processing into mature SFTPB occurs. MVB, multivesicular body.

(B) Western blot of a time course of alveolosphere differentiation (days 0, 16, 22, 29, and 36) and 6-day DCI-cultured primary week 21 human fetal distal lung (HFL DCI-D6) as a positive control. Top: use of an anti-NFLANK antibody that binds to the N-pro region of pro-SFTPB intermediates. Bottom: use of an anti-mature SFTPB antibody (PT3) that binds all SFTPB forms, including the 8-kD mature form.

(C) Dissociation protocol for isolation of intracellular and extracellular components of alveolospheres. Both absolute (nanomoles of lipid/micrograms of DNA in cell samples) and relative (nanomoles of lipid/nanomoles of total diacyl phosphatidylcholine) are shown for both surfactant-specific 32:0 dipalmitoyl phos-phatidylcholine (DPPC) and nonspecific 34:1 phosphatidylcholine (PC). The surfactant index was calculated as the sum of the surfactant-specific 30 and 32 PCs divided by the sum of non-surfactant-specific 36 PCs. Bars represent mean ± SD for each analysis, n = 3 biological replicates separated on day 15; *p % 0.05, **p % 0.01, ***p % 0.001, ****p % 0.0001 by unpaired, two-tailed Student’s t test.