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. 2022 Feb 4;13(2):117. doi: 10.1038/s41419-022-04552-y

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© The Author(s) 2022, corrected publication 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — A WB analysis of changes in CD44, CXCR2, CXCR4, CXCR7, CD74, MIF, pAKT, AKT, pERK1/2, and ERK1/2 expressions in response to IFN-γ (0–500 IU/mL) in A375, SB2, SK-MEL-2, and MeWo. Actin, AKT, and ERK1/2 were used as loading controls. B Release of sCD74 in supernatants 24 h after IFN-γ stimulation (0–500 IU/mL) in A375, SB2, SK-MEL-2, MeWo, THP-1 MΦ, and primary MΦ measured by ELISA (n = 3). C Release of MIF in supernatants 24 h after IFN-γ stimulation (0–500 IU/mL) in A375, SB2, SK-MEL-2, MeWo, THP-1 MΦ, and primary MΦ measured by ELISA (n = 4). D WB analysis of sCD74 in supernatants of A375, SB2, SK-MEL-2, MeWo, and THP-1 MΦ with or without 500 IU/mL IFN-γ stimulation. E WB analysis of sCD74 in the sera of 2 melanoma patients and 2 NHDs. Supernatant of THP-1 MΦ after 500 IU/mL IFN-γ stimulation was used as a reference control. F WB analysis of CD74 in A375 and SK-MEL-2 infected with lentivirus-expressing SC, p33 CD74, and p35 CD74. Parental cells with or without 100 IU/mL IFN-γ stimulation were used as controls. G Release of sCD74 in supernatants under basal conditions in A375 and SK-MEL-2 infected with lentivirus-expressing SC, p33 CD74, and p35 CD74 measured by ELISA (n = 3), and the fold-change relative to sCD74 levels in supernatants of SC cells is shown as bar graphs. H WB analysis of sCD74 in supernatants of A375 and SK-MEL-2 infected with lentivirus-expressing SC, p33, and p35 CD74. Parental cells under 500 IU/mL IFN-γ stimulation were used as a control. I WB analysis of sCD74 in supernatants of A375, SK-MEL-2, and THP-1 MΦ with or without deglycosylation treatment under 500 IU/mL IFN-γ stimulatory conditions. Serum of a melanoma patient was also deglycosylated. J Schematic illustration of deglycosylated full-length p33 CD74^1-216. Considering that MW of deglycosylated sCD74 was approximately 16 KDa, sCD74 was equivalent to a part of full-length CD74 (red box). Graph values represent mean ± SD. CLIP class-II-associated invariant chain peptide, ELISA enzyme-linked immunosorbent assay, IFN-γ interferon-γ, MW molecular weight, MΦ macrophage, NHD normal healthy donor, N.D. not detectable, SC scramble, SD standard deviation, TM transmembrane, WB Western blot.