American as Apple Pie

A vigilant whistleblower sent me this note.

“With Marc Hauser, safely out of the way, faculty at the Ivy League School Harvard may be looking forward to a peaceful and deserved festive season. Or perhaps not? Unfortunately a hot new publication identifying novel E3 ligase substrates in top journal Cell has a number of funnies in the figures.”

Emanuele MJ, Elia AE, Xu Q, Thoma CR, Izhar L, Leng Y, Guo A, Chen YN, Rush J, Hsu PW, Yen HC, Elledge SJ.Global identification of modular cullin-RING ligase substrates.
Cell. 2011 Oct 14;147(2):459-74.
Division of Genetics, Brigham and Women’s Hospital, Boston, MA 02115, USA.

Fig. 6(C) “U2OS cells were transfected with siRNA targeting firefly luciferase (siFF) or the F box proteins Fbxw7 and cyclin F, incubated for 48 hr, and immunoblotted.”

Fig. 7(D) U2OS cells were transfected with siRNA targeting FF, Fbxw7, or cyclin F and, after 48 hr, were treated with UV and harvested 4 hr later for immunoblot.

Loading controls from distinct probes obtained under diverse experimental conditions are not supposed to be identical.

Fig. 6 (F) “U2OS cells were synchronized using a double thymidine block and release. After the first thymidine block, cells were transfected with siRNAs to cyclin F or siFF. Following release, cells were analyzed by immunoblot with the indicated antibodies.
A semiquantitative analysis of NUSAP1 protein levels (relative to loading controls), derived from the western blot shown, is graphed.”

Since loading controls from distinct experiments cannot be identical, the semiquantitative analysis may just as well be false.

Fig. S5(D) “293T, HeLa and U2OS cells were treated with 10 Gy ionizing radiation (IR) and analyzed by immunoblotting of 293T, HeLa and U2OS cells 1, 2 and 4 hr after IR.”

U2OS panel Vinculin appears to be a lighter exposure of 293T.

In order to reject this serious allegation of putative data misrepresentation, the original blots need to be viewed by the authorities concerned.

Figure 1(B) “HeLa and 293T cells were treated for 4 hr with increasing concentrations of MLN4924 (0, 0.1, 1, and 10 mM) and then immunoblotted for NRF2, CDT1, and Vinculin (loading control).”

Hela and 293 NRF2 lanes appear to be duplicates of different exposures. The investigating authorities will have to inspect the original blots to verify.

The whistleblower asks:
“What fate lies in store for this article about cullins? Will it be culled or merely pruned?”

Before making up your mind, though, wait for more intriguing evidence in one of the forthcoming postings.