Journal: Bioscience Reports

Article Title: Inhibition of calpain-1 stabilizes TCF11/Nrf1 but does not affect its activation in response to proteasome inhibition

doi: 10.1042/BSR20180393

Figure Lengend Snippet: ( A and B ) Ea.Hy-926 cells were transfected with siRNA against CAPNS1 or an OFF-Target siRNA. (A) Twenty-four hours later cells were treated with 30 nM BTZ for 16 h, total cell lysates were analysed by immunoblot, representative image from n =3, (B) RNA was isolated and used for qRT-PCR to analyse expression of PSMA3, PSMB6 and PSMC4 , values were calcuated using the ΔΔ C T method with HPRT-1 as housekeeping gene and the normalised to samples without BTZ treatment (set to 1), mean ± SEM, n =3. ( C ) Ea.Hy-926 cells were transfected with flag-tagged constructs for the large subunits of calpain-1 (CAPN1) and -2 (CAPN2). The following day, the cells were treated with 30 nM BTZ for 16 h, total cell lysates were analysed by immunoblot, representative image from n =3.

Article Snippet: The flag-tagged CAPN1 (Addgene #60941) and CAPN2 (Addgene #60942) have been previously described [ ].

Techniques: Transfection, Western Blot, Isolation, Quantitative RT-PCR, Expressing, Construct