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. Author manuscript; available in PMC: 2023 Oct 12.
Published in final edited form as: Cell Rep. 2023 Jun 1;42(6):112577. doi: 10.1016/j.celrep.2023.112577

Figure 2. Telomerase assembly is reduced in the absence of TCAB1.

Figure 2.

(A) Western blots analyzing endogenous TERT immunoprecipitation (IP) probed with a rabbit anti-TERT antibody. TERT level normalized to wild type (WT) (n = 6, mean, SEM).

(B) Northern blot probed for TR and 7SL (loading control) RNA from input and purified endogenous TERT samples. Standards are TR and truncated TRS. Input TR levels relative toWTcontrol normalized to 7SL RNA (n = 3, mean, SD, t test, *p value < 0.05).

(C and D) Quantification of the amount of (C) TR purified relative to input RNA levels, and (D) the ratio of TR relative to endogenous TERT in telomerase purified from TCAB1 knockout cells compared with controls that were normalized to 1 (n = 3, mean, t test).

(E–I) TERT IP from HeLa cells overexpressing TERT and TR (WT) or TRG414C (CABm). (E) Western blots to analyze TERT and TCAB1 levels in cell lysates normalized to total protein levels (n = 3, mean, SD).

(F) Western blots of purified TERT probed with anti-TERT, TCAB1, DKC1, and GAR1 antibodies.

(G) Northern blot probed for TR and 7SL (loading control) of RNA from input and purified endogenous TERT samples. Standards are TR and truncated TRS. Input TR levels relative to WT control normalized to 7SL RNA (n = 3, SD).

(H and I) Quantification of the amount of (H) TR relative to TERT (n = 3, mean, t test), (I) the ratio of TCAB1, DKC1, and GAR1 (n = 3, mean, t test) co-purified with TERT from cells expressing TRG414C (CABm) relative to WT controls normalized to 1.