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closeSupplemental data
Posted by lresar on 09 Jul 2019 at 16:17 GMT
In response to concerns raised by a reader on PubPeer regarding an irregularity in tone in the HMGA1 western blot panel shown in Fig 2D, the authors provide data from repeat western blot experiments carried out to validate the published results. Repeat western blots with 6 biologic replicas (6 tumors/6 matched, non-malignant, adjacent tissue biopsies from the JHU tumor bank) confirm the original results with undetectable HMGA1 in adjacent, non-malignant colon epithelium and high HMGA1 in tumors, with highest levels in invasive tumors. These results are further corroborated at the level of mRNA by qRT-PCR analyses (Fig. 2C), TCGA database analysis by our group in a follow-up study (Xian et al, Nature Commun 2017;8:15008; Fig. 8a), and microarray studies published by another group [Grade et al, Cancer Res 2007;67(1):41-56]. We agree with the reader’s comment that the mark is likely to be a shadow or background mark on the gel or imaging device. The original blots (Fig. 2D) were performed in Dr. Reeves’ laboratory who retired in January 2015 from Washington State University (WSU); WSU is currently retrieving the primary blots from university storage. In accordance with WSU IRB protocol and patient consents, the original patient samples (Fig. 2D) were disposed of when Dr. Reeves retired. Repeat experimental data are now available at https://doi.org/10.6084/m....