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. 2022 Sep 20;41(21):e110372. doi: 10.15252/embj.2021110372

Figure EV2. TSG101 and ATM are equivalently essential for NF‐κB activation by DNA damage.

Figure EV2

  • A
    HEK‐Luc(tGFP) NF‐κB reporter cells were transfected with the same transfection mixtures as used in Fig 2A. Whole‐cell extracts were immunoblotted with antibodies against the indicated proteins. Actin was used as a loading control.
  • B
    U2‐OS cells were transfected with nontargeting (mock) or ATM‐directed siRNAs or three different TSG101‐targeting siRNAs. The indicated cells were treated with etoposide (50 μM, 90 min. Before analysis). Whole‐cell extracts were immunoblotted with the indicated antibodies.
  • C
    U2‐OS cells were transfected with the indicated siRNAs. The NF‐ĸB pathway was activated by etoposide treatment (50 μM, 90 min before analysis). Expression of indicated genes was analyzed using qRT–PCR. The mRNA expression of these genes was normalized to the expression of three housekeeping genes, ACTA1, RPL13A, and TBP2. The gene expression for the indicated conditions is relative to the nontargeting siRNA‐transfected vehicle (DMSO)‐treated cells. The result is representative of three biologically independent experiments. The conditions were compared with an ordinary one‐way ANOVA (*P < 0.05; **P < 0.01). Error bars represent mean ± SEM.