Journal: Open Biology

Article Title: Small molecule protein assembly modulators with pan-cancer therapeutic efficacy

doi: 10.1098/rsob.240210

Figure Lengend Snippet: DRAC of PAV-617 and PAV-51 reveal dynamic multi-protein complex targets. Drug resin affinity chromatography experiments, as described in §4 were carried out with PAV-617 and PAV-951 as well as a negative control resin (134). ( a ) Analysis of PAV-617 resin eluate under hot and cold conditions by silver stain and western blot for PDI, HSP90, KAP-1, YWHAZ, MIF and TUBB with pictures of the western blot underneath the silver stain. The protein band detected by western blot was quantified (graphs of the quantified bands are shown to the right) and statistical significance in different amounts of protein detected in the eluate under different conditions is denoted by asterisks. ( b ) Analysis of PAV-951 resin eluate under hot and cold conditions by silver stain and western blot for PDI, HSP90, KAP-1, ALDH1A1, PSMB5 and GSTP1 with pictures of the western blot underneath the silver stain. The protein band detected by western blot was quantified (graphs of the quantified bands are shown to the right) and statistical significance in different amounts of protein detected in the eluate under different conditions is denoted by asterisks. ( c ) The distribution of the protein ALDH1A1 across a sucrose gradient from starting A549 extract as well as PAV-951 and PAV-621 resin free drug eluate. For each condition, a representative western blot for ALDH1 from triplicate-repeated gradients are pasted below quantitation of the average distribution of the protein across the gradient, with error bars indicating standard deviation. The quantitation shows amount of ALDH1A1 in each particular fraction as a percent of the total ALDH1A1 found across all fractions.

Article Snippet: Drug resin affinity chromatography and SDS–PAGE/silver stain/Western blot analysis of the results, were conducted by Prosetta Biosciences in San Francisco, California, under conditions described in figure legends.

Techniques: Affinity Chromatography, Negative Control, Silver Staining, Western Blot, Quantitation Assay, Standard Deviation