1. BBF2H7-Mediated Sec23A Pathway Is Required for Endoplasmic Reticulum-to-Golgi Trafficking in Dermal Fibroblasts to Promote Collagen Synthesis 
Seiko Ishikura-Kinoshita, Hiroshi Saeki, Kentaro Tsuji-Naito 
Journal of Investigative Dermatology 
Volume 132, Issue 8, Pages (August 2012)
DOI: /jid
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2. Figure 1
IGF-I upregulates the expression of COL1, COL3, and proline 4-hydroxylase (P4H) in normal human dermal fibroblast (NHDF) cultures. NHDF cultures were starved for 24hours, followed by treatment with IGF-I (50ngml−1) for the indicated time. The expression levels of (a) Col1, Col3, and (c) P4h were quantified using real-time PCR. (b, d) Whole-cell lysates were examined by immunoblotting with antibodies for COL1, COL3, P4H, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Values represent mean±SD of triplicate determinations (*P<0.05; **P<0.01 (Tukey–Kramer test)).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
IGF-I upregulates the expression of BBF2 human homolog on chromosome 7 (BBF2H7) and Sec23A. Normal human dermal fibroblast (NHDF) cultures were starved for 24hours, followed by treatment with IGF-I (50ngml−1) or thapsigargin (Tg; 1μM) for the indicated time. The expression levels of (a) Bbf2h7 and (c) Sec23a were quantified using real-time PCR. (b, d) Whole-cell lysates were examined by immunoblotting with antibodies for BBF2H7, Sec23A, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Values represent mean±SD of triplicate determinations (*P<0.05; **P<0.01 (Tukey–Kramer test)).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Bbf2h7 knockdown suppresses the expression of Sec23A and COL1. Normal human dermal fibroblasts (NHDFs) were transfected with the indicated small interfering RNA (siRNA; 20nM) plus HiPerFect Transfection Reagent. At 48hours after transfection, the medium was exchanged for fresh medium with or without IGF-I (50, 100ngml−1) and incubated for 24hours. The expression levels of (a) Sec23a and (b) Col1 were quantified using real-time PCR. (c) Whole-cell lysates were examined by immunoblotting with antibodies for COL1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Values represent mean±SD of triplicate determinations (*P<0.05; **P<0.01 vs. control knockdown and untreated group; ††P<0.01 vs. the indicated group (Tukey–Kramer test)).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
BBF2 human homolog on chromosome 7 (BBF2H7) is induced via the MAPK and phosphoinositide 3-kinase (PI3K) pathways. (a) Normal human dermal fibroblast (NHDF) cultures were starved for 24hours, followed by pretreatment with Tyrphostin AG1024 (10μM) for 30minutes and subsequent treatment with IGF-I (50ngml−1) for 24hours. Whole-cell lysates were examined by immunoblotting with antibodies for COL1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). (b, c) NHDFs cultures were starved for 24hours, followed by pretreatment with Tyrphostin AG1024 (10μM), wortmannin (100nM), or sorafenib (5μM) for 30minutes, and subsequent treatment with IGF-I (50ngml−1) for 8hours. Using real-time PCR, the expression level of Bbf2h7 expression was measured. Values represent mean±SD of triplicate determinations (*P<0.05; **P<0.01 (Tukey-Kramer test)).
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
The BBF2 human homolog on chromosome 7 (BBF2H7)–Sec23A pathway is involved in ER-to-Golgi trafficking. (a–i) Normal human dermal fibroblasts (NHDFs) were transfected with the indicated small interfering RNA (siRNA; 20nM) plus HiPerFect Transfection Reagent. NHDFs were fixed 72hours after transfection and double stained with antibodies for ERGIC-53 and GM130. Bar=10μm. Arrow heads show the irregularly curved Golgi cisternae accompanied by aggregation, characteristic of Golgi dysmorphology. (j) Number of cells showing Golgi dysmorphology per well. NHDFs were cultured at a concentration of 2 × 105cells per well in six-well plates (**P<0.01 (Tukey–Kramer test)).
Journal of Investigative Dermatology  , DOI: ( /jid )
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7. Scheme 1
Proposed mechanisms responsible for collagen synthesis by the BBF2 human homolog on chromosome 7 (BBF2H7)–Sec23A pathway in dermal fibroblasts. ER, endoplasmic reticulum; IGF-IR, IGF-I receptor.
Journal of Investigative Dermatology  , DOI: ( /jid )
Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions