Figure S5.
DSTYK inhibition promotes immunogenic death of lung cancer cells. (A) Top: DSTYK mRNA expression by quantitative PCR in LLC and UN-SCC680 parental and sh#DSTYK cells (n = 3). Bottom: DSTYK protein expression by Western blot in LLC and UN-SCC680 parental and sh#DSTYK cells. Analysis was done by t test. Comparison LLC sh#3 −/+ doxy, P = 0.003; sh#4 −/+ doxy, P < 0.001. Comparison UN-SCC680 sh#3 −/+ doxy and sh#4 −/+ doxy, P < 0.001. (B) Proliferation assay measured by xCELLigence Technology for 96 h in LLC and UN-SCC680 parental and sh#DSTYK cells (n = 3). Analysis was done by t test. (C) DSTYK protein expression by Western blot in parental and sh#DSTYK tumors from LLC and UN-SCC680 preclinical models at the end point of in vivo experiment. (D) Tumor volume of LLC and UN-SCC680 parental or sh#DSTYK cells in immunocompromised mice. Analysis was done by ANOVA. Number of Rag−/− IL2Rg−/− mice per group = 5. (E) T cell killing of LLC parental and KO cell lines was performed in the presence or absence of IFN-γ (n = 3). (F) TNF-α–mediated killing of LLC or H2009 parental, KO, and KO treated with rapamycin cell lines (n = 3). Analysis was performed by t test. Parental vs. KO cells treated with TNF-α, P < 0.001 both in LLC and H2009 cell lines. (G) TNF-α–mediated killing of LLC or H2009 parental, parental treated with chloroquine, and KO cell lines (n = 3). Analysis was performed by t test. Parental vs. parental + chloroquine and parental vs. KO cells treated with TNF-α showed P < 0.001 both in LLC and H2009 cell lines. (H) Cropped images from Western blot analysis of apoptosis and necroptosis-related proteins in H2009 parental and DSTYK-KO cells under normal and stress conditions. (I) Cropped images from Western blot analysis of autophagic protein (LC3) in immortalized 3KT parental and overexpressing DSTYK. (J) T cell killing of 3KT parental and overexpressing DSTYK cell lines was performed in the presence or absence of TNF-α (n = 3). Analysis was performed by t test. P < 0.001 for parental vs. parental treated with TNF-α 3KT cell lines. (K) Tumor volume of parental, KO, and KO overexpressing mutated DSTYK LLC tumor–bearing mice. Analysis was performed by ANOVA. P = 0.002 (parental vs. KO tumor-bearing mice). Number of C57BL/6J mice per group = 7. **, P < 0.01; ***, P < 0.001. Source data are available for this figure: SourceData FS5.

DSTYK inhibition promotes immunogenic death of lung cancer cells . (A) Top: DSTYK mRNA expression by quantitative PCR in LLC and UN-SCC680 parental and sh#DSTYK cells (n = 3). Bottom: DSTYK protein expression by Western blot in LLC and UN-SCC680 parental and sh#DSTYK cells. Analysis was done by t test. Comparison LLC sh#3 −/+ doxy, P = 0.003; sh#4 −/+ doxy, P < 0.001. Comparison UN-SCC680 sh#3 −/+ doxy and sh#4 −/+ doxy, P < 0.001. (B) Proliferation assay measured by xCELLigence Technology for 96 h in LLC and UN-SCC680 parental and sh#DSTYK cells (n = 3). Analysis was done by t test. (C) DSTYK protein expression by Western blot in parental and sh#DSTYK tumors from LLC and UN-SCC680 preclinical models at the end point of in vivo experiment. (D) Tumor volume of LLC and UN-SCC680 parental or sh#DSTYK cells in immunocompromised mice. Analysis was done by ANOVA. Number of Rag−/− IL2Rg−/− mice per group = 5. (E) T cell killing of LLC parental and KO cell lines was performed in the presence or absence of IFN-γ (n = 3). (F) TNF-α–mediated killing of LLC or H2009 parental, KO, and KO treated with rapamycin cell lines (n = 3). Analysis was performed by t test. Parental vs. KO cells treated with TNF-α, P < 0.001 both in LLC and H2009 cell lines. (G) TNF-α–mediated killing of LLC or H2009 parental, parental treated with chloroquine, and KO cell lines (n = 3). Analysis was performed by t test. Parental vs. parental + chloroquine and parental vs. KO cells treated with TNF-α showed P < 0.001 both in LLC and H2009 cell lines. (H) Cropped images from Western blot analysis of apoptosis and necroptosis-related proteins in H2009 parental and DSTYK-KO cells under normal and stress conditions. (I) Cropped images from Western blot analysis of autophagic protein (LC3) in immortalized 3KT parental and overexpressing DSTYK. (J) T cell killing of 3KT parental and overexpressing DSTYK cell lines was performed in the presence or absence of TNF-α (n = 3). Analysis was performed by t test. P < 0.001 for parental vs. parental treated with TNF-α 3KT cell lines. (K) Tumor volume of parental, KO, and KO overexpressing mutated DSTYK LLC tumor–bearing mice. Analysis was performed by ANOVA. P = 0.002 (parental vs. KO tumor-bearing mice). Number of C57BL/6J mice per group = 7. **, P < 0.01; ***, P < 0.001. Source data are available for this figure: SourceData FS5.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal