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. 2022 Feb 5;41(12):1752–1766. doi: 10.1038/s41388-022-02203-2

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© The Author(s) 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. 3 — A Immunoblotting (IB) analysis of Erk3 expression in bone marrow-derived macrophages (BMDM) cultured in the presence (+) or absence (−) of 10% fetal bovine serum. B IB analysis of cell lysates prepared from BMDM treated with different concentrations of etoposide for 24 h. C IB analysis of total cell lysates from the mouse hearts treated with etoposide (50 mg/kg body weight, 24 h). D, E HEK293T cells were transfected different amounts (0–1 µg) of HA-HSC70 or Flag-LAMP2A. Cell lysates were immunoblotted as indicated. F HEK293T and A549 cells were transfected with control siRNA (−) or two sets of Lamp2A siRNAs. Total cell lysates were analyzed for the expression of proteins indicated. G Schematic representation of full-length or different fragments of Erk3 used for the experiments depicted in panels (H) and (I). H IB analysis of cell lysates from HEK293T cells transfected with the indicated plasmids. I IB analysis of cell lysates prepared from HEK293T cells transfected with the indicated plasmids in the presence of Bafilomycin A1 (100 ng/mL, 16 h). J Total cell lysates from HEK293T cells transfected as indicated were immunoprecipitated by anti-Flag and then immunoblotted with anti-HA. K Schematic representation of full-length or different fragments of Erk3 used for the experiments presented in panels (L) and (M). L, M GST pulldown analysis of the interaction between HSC70 and full-length or different fragments of Erk3. N Schematic representation of full-length or different fragments of Erk3 used for experiments depicted in panel (O). O GST pulldown analysis of the interaction between LAMP2A and full-length or different fragments of Erk3. BMDM bone marrow-derived macrophages; FBS fetal bovine serum; WT wild-type; KI knock-in; PC positive control; HSC70 heat shock cognate protein of 70 kDa; LAMP2A lysosome-associated membrane protein type 2A; exp experiment; ctrl. control; IgH heavy chain of IgG; IP immunoprecipitation; IB immunoblotting; PD pull down; GST glutathione S-transferase; siRNA small interfering RNA. Shown are representative images of three independent experiments.