Skip to main content
. 2023 Jun 22;299(8):104948. doi: 10.1016/j.jbc.2023.104948

Figure 2.

Figure 2

Validation by PCR and immunoblotting of a clone with a knock-in biotin tag at the hRpn1 C terminus.A, the binding site of each primer pairing in the PSMD2-edited (left) and PSMD2 parental (right) region is displayed including (1) Biotin-FP1 and 3′-UTR (purple) (2), FP101 and Biotin-RP1 (orange), and (3) FP101 and 3′-UTR (pink) with anticipated amplicon size. The measured amplicon size for each primer set is listed for parental, C1, and C4 in a table to the right. ‘X’ indicates not detected. B–D, representative agarose gel image showing the PCR amplicon generated from (B) primer set 1, (C) primer set 2, and (D) primer set 3, with inclusion of a 1-kilobase-pair (kbp) and 100 base-pair (bp) DNA ladder, as indicated. E, immunoblotting of the whole cell extract from C1 to C6 along with parental WT cells by anti-hRpn1 antibody (left) and HRP-conjugated streptavidin antibody (right). β-actin was also probed as a loading control. The bands corresponding to hRpn1 and hRpn1-biotin are labeled. HRP, horseradish peroxide, UTR, untranslated region.