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[Preprint]. 2023 Feb 14:rs.3.rs-2388864. [Version 1] doi: 10.21203/rs.3.rs-2388864/v1

Figure 2. Knockdown of CXCR4 abrogates trastuzumab resistance.

Figure 2

CXCR4 was silenced by specific shRNA in HCC1419 cells (Materials and Methods). The puromycin-resistant stable colonies were pooled together and named shCXCR4. A pool of cells infected with the lentivirus containing a non-effective vector (shRNA-NE) was selected and used as the control. A Western blot analysis was used to confirm the reduction in CXCR4 expression. B, C CXCR4-knockdown cells or non-silent control cells were co-cultured with BCAFs and PBMCs in 3D, followed by treatment with trastuzumab as illustrated (B). At the endpoint of the study, relative cell viability was quantitatively analyzed using CellTiter-Glo 3D viability assay kit, and the data were analyzed with one-way ANOVA using Prism (C; *P < 0.01, **P < 0.001 compared with the non-silent control cells). D, E CXCR4-knockdown cells or non-silent control cells were used for trastuzumab-induced antibody-dependent cellular cytotoxicity (detail in Materials and Methods). The cells were stained with propidium iodide and analyzed by flow cytometry (D). Data were analyzed using t-test analysis of variance and are reported as the mean ± SD of triplicates (E).