These observations were confirmed by immunohistochemical staining analysis, where elevated expression of Puma, Noxa and Bim, but not Bax proteins could be easily visualised in tumour samples obtained 48 h post-treatment from mice treated with VNB, 48 h before cryotherapy, compared with untreated or single agent-treated mice (Figure 4). Here, PMAIP1 is linked to neoplasm.