HDGF is present in HMCL extracts, HMCL EVs, and HMCL-conditioned media. (A) Top panel, Western blots demonstrate HMCLs express higher levels of HDGF than IVPCs; bottom panel, quantification of HDGF levels of expression normalized to total protein in each sample. Average of 3 experiments normalized to levels in KAS-6/1 (HMCL expressing the highest amount of HDGF). (B) Top panel, Western blots demonstrate EVs derived from HMCLs express varying amounts of HDGF, whereas EVs from IVPCs do not have detectable levels. Bottom panel, quantification of HDGF levels of expression normalized to total protein in each sample. Average of 3 experiments normalized to levels in DP-6 (HMCL EVs expressing the highest amount of HDGF). Nd, not detectable. (C) Levels of HDGF in HMCL-conditioned media measured by ELISA show that HMCLs secrete between 10- to 100-fold more HDGF than IVPCs. HMCL samples were tested in triplicate (except JMW and VP-6, which are duplicates); IVPC values reflect an average of 5 independent determinations. Means ± SD are shown.