Skip to main content
. 2022 Mar 6;18(6):2235–2248. doi: 10.7150/ijbs.64943

Figure 5.

Figure 5

Knockdown of ALKBH5 inhibited the activation of the HIPPO pathway. (A) m6A enrichment of SAV1 was measured by m6A RIP-PCR in ALKBH5- and control RPMI8226 cells. (B) qRT-PCR was used to measure SAV1 mRNA level in the ALKBH5- and control RPMI8226 cells. (C) and (D) mRNA stability of SAV1 in the ALKBH5+ and ALKBH5- RPMI8226 cells was detected by qRT-PCR. (E) Western blot revealed the expression of HIPPO pathway-related proteins, P-YAP (Y357) and P73-target genes (PUMA, P21 and BAX). (F) Western blot revealed the expression of proteins associated with myeloma cell proliferation and apoptosis. (G) RNA immunoprecipitation (RIP) assays in MM RPMI-8226 cells using ALKBH5 and IgG antibody. The ALKBH5-enriched SAV1 mRNA relative to the IgG-enriched value was calculated by qRT-PCR.(I)SAV1 mRNA expression was upregulated after demethylation treatment with DAA in MM RPMI-8226 cells. (H) and (J) Silencing ALKBH5 decreased SAV1 mRNA expression, while DAA treatment increased SAV1 mRNA expression after silencing ALKBH5 in MM cells. Data were mean ± SD. *P<0.5, **** P<0.0001.