Journal: Communications Biology

Article Title: Arabidopsis LSH10 transcription factor and OTLD1 histone deubiquitinase interact and transcriptionally regulate the same target genes

doi: 10.1038/s42003-023-04424-x

Figure Lengend Snippet: a Association of LSH10-His6 with the chromatin of the indicated target genes in the lsh10-1/LSH10-His6 plants. lsh10-1/LSH10-His6 , white bars; non-specific background signal obtained in the wild-type plants that do not express the His6 epitope, gray bars. The outside probe is the qChIP probe, depicted in Fig. , and located outside the EMSA probe locations. b Increase in H2B monoubiquitylation of the OSR2, WUS, ABI5 , and ARL promoter chromatin in the lsh10-1 mutant plants. Wild-type plants, dark gray bars; lsh10-1 , light gray bars; lsh10-2 , white bars. The chromatin association of LSH10-His6 and the degree of H2B monoubiquitylation were analyzed by qChIP with probes described in Fig. and listed in Supplementary Data . Error bars represent the SEM of n = 5 or n = 7 biological replicates with 3 technical repeats for each. The individual data points are indicated, and their numerical values are listed in Supplementary Data and Supplementary Data . Differences between mean values assessed by the two-tailed t-test are statistically significant for the p -values * p < 0.05 and ** p < 0.01; P ≥ 0.05 are not statistically significant (ns).

Article Snippet: 5 µl of the crude chromatin extracts were saved for use as an input control, and 100 µl were added into the microwell immobilized with the antibodies (non-specific rabbit IgG Isotype Control (Invitrogen, WB317638, 1 µg) as the negative control, specific rabbit anti-His-tag antibody (GenScript, A00174-40, 2.5 µg) or anti-monoubiquityl-histone H2B (Lys-120) (5546S, Cell Signaling Technology, Inc., 2.5 µg).

Techniques: Mutagenesis, Two Tailed Test