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. 2024 Feb 6;84(9):1475–1490. doi: 10.1158/0008-5472.CAN-23-2812

Figure 4.

Figure 4. Targeting TACC3 sensitizes the human HER2-expressing EMT6.huHER2 cells to T-DM1 and induces ICD markers. A, Cell viability assay in EMT6.huHER2 cells treated with increasing doses of T-DM1 alone or combination with different doses of BO-264 for 3 days (n = 4). B, Validation of TACC3 knockout in EMT6.huHER2 cells obtained using CRISPR/Cas9 system. C, Cell viability assay in EMT6.huHER2.sgTACC3 vs. sgControl cells treated with increasing doses of T-DM1 for 3 days (n = 4). D, Western blot analysis of mitotic arrest, apoptosis, and ICD markers in EMT6.huHER2 cells treated with T-DM1 alone or in combination with BO-264. Actin was used as a loading control. E, Western blot analysis of TACC3, mitotic arrest, apoptosis, and ICD markers in EMT6.huHER2.sgTACC3 vs. sgControl cells treated with T-DM1. Actin was used as a loading control. F, Relative ATP release from EMT6.huHER2 cells treated with T-DM1 alone or in combination with BO-264 (n = 3). G, Relative ATP release from EMT6.huHER2.sgTACC3 vs. sgControl cells treated with T-DM1 (n = 3, 4). H, Immunofluorescence cell-surface staining of calreticulin (green) in EMT6.huHER2 cells treated with T-DM1 alone or in combination with BO-264. Its quantification is provided on the right. I, Immunofluorescence cell-surface staining of calreticulin (green) in EMT6.huHER2.sgTACC3 vs. sgControl cells treated with T-DM1. Its quantification is provided on the right. Data correspond to mean values ± SD. P values were calculated with the unpaired, two-tailed Student t test. **, P < 0.01.

Targeting TACC3 sensitizes the human HER2-expressing EMT6.huHER2 cells to T-DM1 and induces ICD markers. A, Cell viability assay in EMT6.huHER2 cells treated with increasing doses of T-DM1 alone or combination with different doses of BO-264 for 3 days (n = 4). B, Validation of TACC3 knockout in EMT6.huHER2 cells obtained using CRISPR/Cas9 system. C, Cell viability assay in EMT6.huHER2.sgTACC3 vs. sgControl cells treated with increasing doses of T-DM1 for 3 days (n = 4). D, Western blot analysis of mitotic arrest, apoptosis, and ICD markers in EMT6.huHER2 cells treated with T-DM1 alone or in combination with BO-264. Actin was used as a loading control. E, Western blot analysis of TACC3, mitotic arrest, apoptosis, and ICD markers in EMT6.huHER2.sgTACC3 vs. sgControl cells treated with T-DM1. Actin was used as a loading control. F, Relative ATP release from EMT6.huHER2 cells treated with T-DM1 alone or in combination with BO-264 (n = 3). G, Relative ATP release from EMT6.huHER2.sgTACC3 vs. sgControl cells treated with T-DM1 (n = 3, 4). H, Immunofluorescence cell-surface staining of calreticulin (green) in EMT6.huHER2 cells treated with T-DM1 alone or in combination with BO-264. Its quantification is provided on the right. I, Immunofluorescence cell-surface staining of calreticulin (green) in EMT6.huHER2.sgTACC3 vs. sgControl cells treated with T-DM1. Its quantification is provided on the right. Data correspond to mean values ± SD. P values were calculated with the unpaired, two-tailed Student t test. **, P < 0.01.