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a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
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cd25 apc  (Miltenyi Biotec)
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Miltenyi Biotec cd25 apc
a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
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pe antihuman foxp3  (Miltenyi Biotec)
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a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
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cd25  (Miltenyi Biotec)
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Miltenyi Biotec cd25
a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
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cd25 microbeads ii  (Miltenyi Biotec)
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a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
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av142  (Bio-Rad)
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Bio-Rad av142
a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
Av142, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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cd25 selection  (Miltenyi Biotec)
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Miltenyi Biotec cd25 selection
a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
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mouse anti chicken cd25  (Bio-Rad)
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Bio-Rad mouse anti chicken cd25
a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
Mouse Anti Chicken Cd25, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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reafinitytm  (Miltenyi Biotec)
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a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs <t>(CD4+CD25+)</t> of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .
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Image Search Results


a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs (CD4+CD25+) of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .

Journal: Nature Communications

Article Title: Nrf2-mediated metabolic reprogramming drives regulatory T cell accumulation in hepatocellular carcinoma

doi: 10.1038/s41467-026-73485-3

Figure Lengend Snippet: a Representative gating strategy of Nrf2-activated cells (HO-1+) within Tregs (CD127 lo CD25 hi FoxP3+) and CD4+ Tconv (CD127+ CD25−). b , c Frequency of live and Annexin V+ cells within HO-1+ or HO-1-Tregs and CD4+ Tconv. d – i Frequency of live and Annexin V+ cells in donor Tregs (CD4+CD127 lo CD25 hi ) and Tconv (CD4+ CD25− and CD8+) before (baseline) and after 24 h culture in RPMI standard or high lactate and low glucose with and without the NRF2 inhibitor ML385. j , k Frequency of live and Annexin V+ cells of Tregs (CD4+ CD25+) and Teff (CD4+ CD25− and CD8+) of WT and Nrf2 −/− mice after 24 h culture in RPMI standard or high lactate and low glucose. l Example histograms of the mean fluorescence intensity (MFI) of MtGreen and MtDR in freshly isolated Tregs (CD4+CD25+) of WT and Nrf2 −/− mice. m Ratio of the geometric MFI (gMFI) of MtGreen and MtDR and n radiometric index on freshly isolated Tregs, Tconv (CD4+ CD25− or CD8+) of WT and Nrf2 −/− mice. The radiometric index was calculated as the difference between the log₁₀-transformed MFI of MtDR and the log₁₀-transformed MFI of MtGreen. o Example histogram on Tregs and CD4+ Tconv (left, CD4+CD25+ and CD4+ CD25−) and mean MFI (right) of MtGreen and MtDR in HCC samples. Data are presented as mean values + SD or ±SD as appropriate. P -values were calculated by two-way ANOVA followed by Tukey’s multiple comparisons post-hoc test for ( b – n ); or paired t-test (two-sided) for ( o ). Only statistically significant P -values are displayed: *, P < 0.05; **, P < 0.01; ***, P < 0.001. MtDR, mitotracker deep red; MtGreen, mitotracker green. FACS sequential gating strategy for ( d – k ) is provided in Supplementary Fig. .

Article Snippet: Freshly isolated human healthy CD4+CD25+ Tregs were pre-incubated in the presence or absence of 100 nM of the MCT1 inhibitor AZD3965 (MedChemExpress; HY-12750) for 2 h and then cultured with low dose IL-2 (50IU/ml) and 1:5 aCD3/CD28 beads in RPMI standard or RPMI high lactate and low glucose (as described above) for 30 min.

Techniques: Fluorescence, Isolation, Transformation Assay