a300 362a Search Results


cbp  (Fortis Life Sciences)
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Fortis Life Sciences cbp
Reported HAT inhibitors cause nonspecific cell assay readouts. a Double p300 and <t>CBP</t> knockdowns do not affect cell viability after 2 d. Data are expressed as mean ± SEM of three technical replicates (ns, not significant; two-tailed Student’s t -test). b Double p300 and CBP knockdowns efficiently <t>reduce</t> <t>H3K27ac</t> levels in HEK293T and MCF7 cells. Molecular weight of probed protein is indicated in kilodaltons (kDa) as verified by molecular weight markers. Data are representative results from one of two independent experiments. c Many reported HAT inhibitors exert anti-proliferative effects at low micromolar compound concentrations in (top) MCF7 and (bottom) HEK293T cells as monitored by cell confluence 24 h after addition of compound. Note the nonspecific interference compounds NSC-663284 (N), rottlerin (R), and 24 – 27 (chemical structures shown as inset) demonstrate similar anti-proliferative effects. C, DMSO control. Red error bars, statistically significant difference from respective DMSO controls ( p < 0.05; two-tailed Student’s t -test and Holm–Sidak method). Data are expressed as mean ± SD of three technical replicates and are representative results from one of two independent experiments. d Many reported HAT inhibitors also cause nonspecific changes in H3K27ac and p300 levels in cells at low micromolar compound concentrations. Molecular weights of protein analytes are indicated in kDa as verified by molecular weight markers (Supplementary Fig. ). Note the same known interference compounds from panel c can also decrease H3K27ac levels. Data are representative results from one of two independent experiments
Cbp, supplied by Fortis Life Sciences, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Reported HAT inhibitors cause nonspecific cell assay readouts. a Double p300 and CBP knockdowns do not affect cell viability after 2 d. Data are expressed as mean ± SEM of three technical replicates (ns, not significant; two-tailed Student’s t -test). b Double p300 and CBP knockdowns efficiently reduce H3K27ac levels in HEK293T and MCF7 cells. Molecular weight of probed protein is indicated in kilodaltons (kDa) as verified by molecular weight markers. Data are representative results from one of two independent experiments. c Many reported HAT inhibitors exert anti-proliferative effects at low micromolar compound concentrations in (top) MCF7 and (bottom) HEK293T cells as monitored by cell confluence 24 h after addition of compound. Note the nonspecific interference compounds NSC-663284 (N), rottlerin (R), and 24 – 27 (chemical structures shown as inset) demonstrate similar anti-proliferative effects. C, DMSO control. Red error bars, statistically significant difference from respective DMSO controls ( p < 0.05; two-tailed Student’s t -test and Holm–Sidak method). Data are expressed as mean ± SD of three technical replicates and are representative results from one of two independent experiments. d Many reported HAT inhibitors also cause nonspecific changes in H3K27ac and p300 levels in cells at low micromolar compound concentrations. Molecular weights of protein analytes are indicated in kDa as verified by molecular weight markers (Supplementary Fig. ). Note the same known interference compounds from panel c can also decrease H3K27ac levels. Data are representative results from one of two independent experiments

Journal: Nature Communications

Article Title: Assay interference and off-target liabilities of reported histone acetyltransferase inhibitors

doi: 10.1038/s41467-017-01657-3

Figure Lengend Snippet: Reported HAT inhibitors cause nonspecific cell assay readouts. a Double p300 and CBP knockdowns do not affect cell viability after 2 d. Data are expressed as mean ± SEM of three technical replicates (ns, not significant; two-tailed Student’s t -test). b Double p300 and CBP knockdowns efficiently reduce H3K27ac levels in HEK293T and MCF7 cells. Molecular weight of probed protein is indicated in kilodaltons (kDa) as verified by molecular weight markers. Data are representative results from one of two independent experiments. c Many reported HAT inhibitors exert anti-proliferative effects at low micromolar compound concentrations in (top) MCF7 and (bottom) HEK293T cells as monitored by cell confluence 24 h after addition of compound. Note the nonspecific interference compounds NSC-663284 (N), rottlerin (R), and 24 – 27 (chemical structures shown as inset) demonstrate similar anti-proliferative effects. C, DMSO control. Red error bars, statistically significant difference from respective DMSO controls ( p < 0.05; two-tailed Student’s t -test and Holm–Sidak method). Data are expressed as mean ± SD of three technical replicates and are representative results from one of two independent experiments. d Many reported HAT inhibitors also cause nonspecific changes in H3K27ac and p300 levels in cells at low micromolar compound concentrations. Molecular weights of protein analytes are indicated in kDa as verified by molecular weight markers (Supplementary Fig. ). Note the same known interference compounds from panel c can also decrease H3K27ac levels. Data are representative results from one of two independent experiments

Article Snippet: Blots were blocked for 1 h in blocking buffer (5% milk in 0.1% Tween 20/PBS) and incubated with respective primary antibodies: H3K27ac (1:1000, CST #8173), H3 (1:1000, Abcam #10799), p300 (1:2000, Bethyl #A300-358A), CBP (1:1000, Bethyl #A300-362A), actin (1:3000, Abcam #3280), α-tubulin (1:1000, 12G10 DHSB), and α-tubulin-K40ac (1:2000, Abcam #179484) in blocking buffer overnight at 4 °C.

Techniques: Two Tailed Test, Molecular Weight