Global proteomics of HEK293T cells treated with Val-boroPro for 48h

To find cellular protein binding partners of the NLRP1 inflammasome, FLAG-tagged full length NLRP1 (or a GFP-FLAG protein control) was ectopically expressed in HEK 293T cells and affinity purified from cell lysates (Note: the NLRP1-FLAG-containing lysate samples were affinity purified in the presence or absence of excess proline) before proteins were reduced, alkylated, and trypsinized before TMT...

HEK293T Azidohomoalanine (AHA) Pulse Chase (degradome) time course experiment, were cells from various genetic background (WT, ATG7-/-, RB1CC1-/-) are treated or not with Torin1 for 5h, 10h or 15h. Samples are Biotin-click, and streptavidin enriched, digested and labeled with TMTpro 16plex.

HEK293T subject to Torin1 treatment or amino acid withdraw dataset and translatome analysis was determined using Azidohomoalanine (AHA) incorportation, Biotin-click, and streptavidin enrichment. TMT 10plex labelling was used. Three genotypes, WT (24 fractions), ATG7 KO (HA-9-53-1 single shot), RB1CC1 KO (HA-9-53-2 single shot).

HEK293 cells with two genotype (WT or ATCG5 KO), treated with Torin1 or amino acid withdraw. TMT 11 plex.

Target identification of methyl bestatin (MeBs) using the photoreactive diazirine-alkyne probe, CQ83.

HEK293T proteome (single shot) with knock-in or not for RPS3-Halo and Knock out or not for NUFIP1. Cells were treated or not with Torin1 or deprived of amino acids.

HEK293T subject to Single AA starvation dataset and translatome analysis was determined using Azidohomoalanine (AHA) incorportation, Biotin-click, and streptavidin enrichment. TMTpro 16plex labels was used.

HEK293T cells were treated with 6-MP and harvested at various time point (6, 9 and 18h). Translatome was measure by Azidohomoalanine (AHA) incorporation, Biotin-click, and streptavidin enrichment.

Summary from GEO: "A canonical pathway generates microRNAs (miRNAs) via stepwise cleavage of hairpin precursors by RNase III enzymes Drosha and Dicer, followed by loading of resultant ~22 nucleotide (nt) RNAs into an Argonaute (Ago) effector complex. In addition, non-canonical substrates can bypass either RNase III enzyme to yield functional small RNAs, including hundreds of intron-derived hairpins...