Journal: Clinical cancer research : an official journal of the American Association for Cancer Research
Article Title: A Novel Splice Variant of HYAL-4 Drives Malignant Transformation and Predicts Outcome in Bladder Cancer Patients.
doi: 10.1158/1078-0432.CCR-19-2912
Figure Lengend Snippet: A: Immunoblot analysis of bladder tissue extracts using anti-HYAL-4 antibody. B: Chase levels (U/mg protein) in bladder tissue extracts measured by Chase assay. C: HYAL-4 staining in representative NBL and BC tissues. Magnification: 400X. See Figure S2 for enlarged photos and quantification. D: Immunoblot anlaysis of cell lysates and CM of EV, Wt, and V1, Ctr, and HYAL-4 shRNA transfectants for indicated proteins; loading control: Actin. HYAL-4 peptide block: the peptide was added during incubation with the anti-HYAL-4 antibody. Note for A and D: In each blot, all sample lanes were run on the same gel with the same exposure time; a gap denotes those samples that were not contiguous within the gel. E: Chase activity (U) was measured in the CM of bladder cell transfectants (left panel) and in the CM of control (Ctr) and HYAL-4 shRNA transfectants (right panel). The activity was normalized to cell number. Data: Mean ± SEM; quadruplicate. F: Normalized Chase activity in the CM of HT1376 transfectants and in a high-grade bladder tumor tissue extract (HG-TBL) at indicated pH. Data: Mean ± SEM; quadruplicate. G: Chase activity in the CM of HT1376 V1 transfectants and a HG-TBL extract measured against different chondriotin sulfate substrates. Data: Mean ± SEM; quadruplicate.
Article Snippet: HYAL-4 shRNA transfectants: HYAL-4 shRNA constructs were purchased from Origene.
Techniques: Western Blot, Staining, shRNA, Blocking Assay, Incubation, Activity Assay