Figure 2.

BRD9 inhibition abolishes CSC characteristics and blocks cells from entering the G0 phase. (A) BRD9 inhibition sensitizes CSCs for gemcitabine-mediated elimination. (B and C) Triple-positive OCT4−GFP+/CD133+/SSEA4+ CSC marker reduction in FG cells is induced by gemcitabine cotreatment with I-BRD9 or SB431542 as shown in bar graph (B) or density maps from flow cytometry analyses (C). (D) TGFβ/Activin signaling inhibition and BRD9 inhibition impair CSC self-renewal and sensitize CSCs for gemcitabine-mediated destruction. (E) BRD9 inhibition shifts the balance between CSCs and non-CSCs toward non-CSCs. (F) Schematic depiction of the 3-color cell cycle analysis system. (G) Fluorescence microscopy of 3FUCCI-PDAC cells grown in 2D condition. (H) Fluorescence microscopy overlayed with bright field image of 3FUCCI-PDAC cells grown in 3D sphere condition. (I) BRD9 inhibition blocks cells from entering the G0 phase. (J and K) BRD9 inhibition reduces cell migration. (J) Normalised data of wound-healing assay as a bar graph. (L and M) BRD9 inhibition reduced PDAC cell invasiveness. (N) Schematic overview of the SMAD2/3 proteomic experiment. (O) SS18/SSXT, BAF180/Polybromo-1, and ARID1A form a complex with SMAD2/3 in CSCs. (P) SMAD2/3 proteins interact with the subunits of noncanonical BAF, esBAF, and npBAF complexes in CSCs. (R) TGFβ/Activin A signaling leads to the formation of protein complex between SMAD2/3 and BRD9/BAF that regulates the function of chemoresistance, G0 phase entry, migration, and invasiveness of CSCs. Experiments represent 3 replicates. Statistical analysis was performed by 2-way analysis of variance with multiple comparisons with Tukey correction. ****Adjusted P < .0001, ***adjusted P < .001, **adjusted P < .01, *adjusted P < .05.