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anti nrf2  (Proteintech)
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Proteintech anti nrf2
( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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nfe2l2  (Proteintech)
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .
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( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .

Journal: Science Advances

Article Title: NRF2 coordinates cystine import and glutathione use to sustain ferroptosis resistance and formaldehyde tolerance in cancer

doi: 10.1126/sciadv.aea1431

Figure Lengend Snippet: ( A ) Overview of cytosolic formaldehyde (FA) catabolism. ( B ) Heatmap depicting Log10 of the 50% FA inhibitory concentration (IC 50 ) in the presence or absence of l -buthionine-sulfoximine (BSO) at concentrations of 25 μM (H2170 cells), 50 μM (Jurkat, Nalm6, K562, and LUDLU-1 cells) or 100 μM (HCT116, H1299, A549, and T98G cells). ( C ) Representative FA viability curves normalized (norm.) to 0 μM FA for H1299 and A549 cells [mean ± SEM, n = 5 (H1299), n = 6 (A549), and two-way ANOVA (Tukey test between untreated (UT) and 100 μM BSO); for H1299: ** P = 0.0016, **** P < 0.0001, and ** P = 0.0023]. ( D ) Glutathione (GSH) levels upon 24-hour exposure to BSO [same concentrations as (B)] (mean ± SEM, n represented as dots, t test comparison between UT and BSO; * P = 0.0142, ** P = 0.0073, **** P < 0.0001). ( E ) Colony survival assay (CSA) in HCT116, H1299, and A549 in the presence of FA and/or 3.12 μM BSO for H1299 cells and 6.25 μM BSO for both HCT116 and A549 cells [mean ± SEM, n represented as dots, one-way ANOVA (Tukey test); for HCT116: **** P < 0.0001; for H1299: * P = 0.0106 and ** P = 0.0033]. ( F ) Left: GSH quantification in A549 cells expressing a nontargeting-control sgRNA (sgNTC) or sgRNAs against GSS (sg GSS ). Right: Viability in response to FA [mean ± SEM, n = 3]. ( G ) Immunoblots in A549 cells carrying an empty vector (pEV) or a vector coding for wild-type KEAP1 (p KEAP1 ). ( H ) Viability curves for A549/pEV or A549/pKEAP1 cells exposed to different concentrations of FA in the presence of 100 μM BSO [mean ± SEM, n = 3 and two-way ANOVA (Tukey test between UT and 100 μM BSO); **** P < 0.0001]. ( I ) DNA-protein cross-link (DPC) accumulation in A549/pEV and A549/pKEAP1 cells exposed to 100 μM BSO for 48 hours and to the indicated concentrations of FA for 4 hours (mean ± SEM, n = 3, one-way ANOVA in each cell line corrected by Tukey test) . Created in BioRender. https://BioRender.com/tsuwhsf .

Article Snippet: The following antibodies were used across this work: Anti-SLC7A11/xCT (Abcam, catalog no. ab307601; RRID: AB_3094570), Anti-GCLC (H-5) (Santa Cruz Biotechnology, catalog no. sc-390811; RRID: AB_2736837), Anti-GCLM (G-4) (Santa Cruz Biotechnology, catalog no. sc-55585; RRID: AB_831788), Anti-ADH5 (2D11) (Santa Cruz Biotechnology, catalog no. sc-293460; RRID: AB_2927735), Anti–α-Tubulin (DM1A) (Santa Cruz Biotechnology catalog no. sc-32293 HRP; RRID: AB_628412), Anti-EIF2S1 (A42:F421) (Proteintech, catalog no. 11170-1-AP; RRID: AB_2096489), Anti–p-EIF2S1 (Ser51) (Proteintech, catalog no. 68023-1-Ig; RRID: AB_2918767), Anti-GSS (OTI2F2) (OriGene, catalog no. TA501936; RRID: AB_11128188), Anti-NRF2 (Proteintech, catalog no. 16396-1-AP; RRID: AB_2782956), Anti-Vinculin (7F9) (Santa Cruz Biotechnology, catalog no. sc-73614 HRP; RRID: AB_1131294), Anti-GAPDH (6C5) (Abcam, catalog no. ab8245; RRID: AB_2107448), Anti-Puromycin (12D10) (Merck, catalog no. MABE343; RRID: AB_2566826), Anti-FLAG (M2) (Merck, catalog no. F3165; RRID: AB_259529), IRDye 680RD Goat anti-Ms (LI-COR, catalog no. 926-68070; RRID: AB_10956588), IRDye 800CW Donkey anti-Rb (LI-COR, catalog no. 926-32213; RRID: AB_621848), Anti-Rabbit IgG-HRP (Merck, catalog no. A0545; RRID: AB_257896), and Anti-Mouse IgG-HRP (Merck, catalog no. A9044; RRID: AB_2313567).

Techniques: Concentration Assay, Comparison, Clonogenic Cell Survival Assay, Expressing, Control, Western Blot, Plasmid Preparation