Journal: Journal of Biological Chemistry

Article Title: Lanthionine Synthetase C-like Protein 1 Interacts with and Inhibits Cystathionine β-Synthase

doi: 10.1074/jbc.m112.383646

Figure Lengend Snippet: FIGURE 7. LanCL1 knockdown increases CBS activity and protects neu- rons against H2O2 or glutamate. A, left, shown are representative immuno- blots of LanCL1 and CBS in neurons infected with LanCL1 RNAi or control RNAi lentivirus. LanCL1 RNAi decreased the expression of LanCL1. Right, shown is quantitative analysis of three independent experiments. The ratio of the band intensity of LanCL1 to actin in control neurons (Control RNAi) was normalized to 100% (*, p 0.05; unpaired t test; n 3). B, lentiviral knock- down of LanCL1 increased the CBS activity in neurons under basal conditions or with H2O2 or glutamate exposure. LanCL1 RNAi occluded the H2O2 or glu- tamate-induced increase of CBS activity (*, p 0.05 versus control RNAi; n.s., no significant difference versus basal; unpaired t test; n 5). C, for cell viability measured by methylthiazolyldiphenyl-tetrazolium bromide assay, the values were normalized to the respective controls (defined as 100%). LanCL1 knock- down increased neuronal viability after H2O2 or glutamate exposure for 2 h and another 24 h in culture (*, p 0.05; unpaired t test; n 3). D, for apoptosis analyzedbyHoechststaining,theratioofthenumberofapoptoticneuronsto the total number was normalized to the respective control (defined as 100%). LanCL1 knockdown decreased the proportion of apoptotic neurons after H2O2 or glutamate challenge (*, p 0.05; unpaired t test; n 3). Data are mean S.E.

Article Snippet: Lentivirus-mediated RNAi in Primary Rat Cortical Neurons— LanCL1 shRNA (rat) lentiviral particles (SC-270327-V) and control shRNA lentiviral particles were from Santa Cruz.

Techniques: Knockdown, Activity Assay, Western Blot, Infection, Control, Expressing

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: Clinical characteristics of PCa patients and LanCL1 expression

Article Snippet: The primary antibody was LanCL1 (Proteintech, 1/600, 30 min).

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a , b LanCL1 overexpression in LNCaP and PC-3 cells reduced cell death after 24 h treatment of 150 μM H2O2. Student’s t -test was performed for statistical significance analysis. N = 5. c , d LanCL1 knockdown increased LNCaP and PC-3 cells death induced by H 2 O 2 (100 μM H 2 O 2 , 24 h treatment). Student’s t -test was performed for statistical significance analysis. N = 5. e Hoechst staining shows that downregulation of LanCL1 increased LNCaP cell death(indicated by arrows) induced by H 2 O 2 , while LanCL1 overexpression reduced cell death. f Quantitation of the Hoechst staining of LNCaP cell treated by H 2 O 2 . Student’s t -test was performed for statistical significance analysis. N >3

Article Snippet: The primary antibody was LanCL1 (Proteintech, 1/600, 30 min).

Techniques: Over Expression, Knockdown, Staining, Quantitation Assay

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues. Prostate is indicated by red arrow. b IHC data from The Human Protein Atlas portal show LanCL1 expression is higher in prostate cancer tissues than in normal tissues. c . Western blotting was used to detect the LanCL1 expression in human prostate cell lines. Tubulin served as the control. d Quantitation of relative expression of LanCL1 protein in different prostate cancer cell lines. e , f Increased expression markedly correlates with high Gleason score and tumor stage, data from the TCGA data set (queried from cBioPortal). LanCL1 expression is exon-normalized signal intensity. Mann–Whitney U -tests were performed to assess statistical significance for the comparisons between groups

Article Snippet: The primary antibody was LanCL1 (Proteintech, 1/600, 30 min).

Techniques: Expressing, Western Blot, Control, Quantitation Assay, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a Representative immunohistochemistry of LanCL1 on benign prostatic epithelia (benign) and prostate cancer tissues (Gleason score 6 or 9). Student’s t-test was performed for statistical significance analysis. b Quantitation of protein expression of LanCL1 in benign or prostate cancer tissues. c Western blot analysis of LanCL1 expression in 15 pairs of nontumor tissues (P) and PCa tissues (T). Student’s t-test was performed for statistical significance analysis. N = 15. d Immunohistochemical staining for LanCL1 protein in the dorsolateral prostate of WT and TRAMP mice

Article Snippet: The primary antibody was LanCL1 (Proteintech, 1/600, 30 min).

Techniques: Immunohistochemistry, Quantitation Assay, Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a Stably transfected LNCaP cells and PC-3 cells interfered with LanCL1 were established. LanCL1 was knockdown by siRNA as indicated. The efficiency of knockdown were examined by western blotting. Tubulin served as the control. b Cell proliferation was measured at the indicated time points by MTS colorimetric assay (absorbance at 490 nm (OD490). Student’s t -test was performed for statistical significance analysis. N >3. * P <0.05, ** P <0.01, *** P <0.001 c Tumor growth in LanCL1 overexpressed PC-3 cells and control cells was investigated by xenograft tumor models. Student’s t-test was performed for statistical significance analysis. N = 7. ** P <0.01, *** P <0.001. d The cell cycle distribution was analyzed by fluorescent-activated cell scanning (FACS) in LanCL1 overexpressed LNCaP cells. e Column chart indicates the mean and SEM of the S-phase fraction for each cell. Student’s t-test was performed for statistical significance analysis. N = 5

Article Snippet: The primary antibody was LanCL1 (Proteintech, 1/600, 30 min).

Techniques: Stable Transfection, Transfection, Knockdown, Western Blot, Control, Colorimetric Assay

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a qRT–PCR shows no induction of LanCL1 mRNA in H 2 O 2 -treated (100 μM, 1 h) LNCaP cells. Student’s t -test was performed for statistical significance analysis. N = 3. b , c Western blots and quantification show that H 2 O 2 -treatment did not induce LanCL1 protein expression. Student’s t-test was performed for statistical significance analysis. N = 5. d – f Western blots and quantification show an increase in the level of 4-HNE in the H 2 O 2 -treated (100 μM, 1 h) LNCaP cells, but LanCL1 overexpression did not influence the level of 4-HNE. Western blots show that LanCL1 overexpression in LNCaP cells is not more resistant to H 2 O 2 -induced 4-HNE accumulation. Student’s t-test was performed for statistical significance analysis. N = 5. g ROS determination by using a fluorescent probe DCFH-DA in LNCaP and PC-3 cells untreated and treated with H 2 O 2 (100 μM, 1 h). Student’s t-test was performed for statistical significance analysis. N = 3

Article Snippet: The primary antibody was LanCL1 (Proteintech, 1/600, 30 min).

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Over Expression

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a , b Modulation of apoptosis related signaling proteins using antibody array assay. Total protein was extracted from LNCaP-NEO and LNCaP-LanCL1 cells. A slide-based antibody array was used for simultaneous detection of 19 signaling molecules involved in stress response and apoptosis using a PathScan Stress and Apoptosis Signaling Array kit. Each protein was arranged in duplicate. c The proteins with changes concentration included pP44/42 MAPK (ERK1/2) (Thr202/Try204), pAkt(S473), pHSP27(Ser82), pSmad2(Ser465), pP53(Ser15), pP38 MAPK(Thr180/Try182), pSAPK/JNK(Thr183/Try185) and Cleaved-Caspase-3(Asp175), which were indicated on the images, and quantified. d The concentrations of the protein in c were quantified. Student’s t -test was performed for statistical significance analysis. N = 3. Data are presented as mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001. e Luciferase assay shows that overexpression of LanCL1 reduced AP-1 transcription. Student’s t-test was performed for statistical significance analysis. N = 3. *** P <0.001 f Western blotting indicated the protein level of pSAPK/JNK(Thr183/Try185) in overexpression LNCaP and PC-3 cells. Student’s t -test was performed for statistical significance analysis. N = 3. g Western blotting indicated the protein level of pSAPK/JNK(Thr183/Try185) in overexpression LNCaP cells after 100 μM H 2 O 2 for 2 h. Student’s t-test was performed for statistical significance analysis. N = 3. H. SP600125 partially rescued the cell death caused by ROS in LanCL1 knockdown LNCaP cells. Student’s t-test was performed for statistical significance analysis. N = 5

Article Snippet: The primary antibody was LanCL1 (Proteintech, 1/600, 30 min).

Techniques: Ab Array, Concentration Assay, Luciferase, Over Expression, Western Blot, Knockdown

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: Clinical characteristics of PCa patients and LanCL1 expression

Article Snippet: Fig. 2 Associations between LanCL1 expression and prostate cancer risk and severity. a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues.

Techniques: Expressing

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a , b LanCL1 overexpression in LNCaP and PC-3 cells reduced cell death after 24 h treatment of 150 μM H2O2. Student’s t -test was performed for statistical significance analysis. N = 5. c , d LanCL1 knockdown increased LNCaP and PC-3 cells death induced by H 2 O 2 (100 μM H 2 O 2 , 24 h treatment). Student’s t -test was performed for statistical significance analysis. N = 5. e Hoechst staining shows that downregulation of LanCL1 increased LNCaP cell death(indicated by arrows) induced by H 2 O 2 , while LanCL1 overexpression reduced cell death. f Quantitation of the Hoechst staining of LNCaP cell treated by H 2 O 2 . Student’s t -test was performed for statistical significance analysis. N >3

Article Snippet: Fig. 2 Associations between LanCL1 expression and prostate cancer risk and severity. a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues.

Techniques: Over Expression, Knockdown, Staining, Quantitation Assay

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues. Prostate is indicated by red arrow. b IHC data from The Human Protein Atlas portal show LanCL1 expression is higher in prostate cancer tissues than in normal tissues. c . Western blotting was used to detect the LanCL1 expression in human prostate cell lines. Tubulin served as the control. d Quantitation of relative expression of LanCL1 protein in different prostate cancer cell lines. e , f Increased expression markedly correlates with high Gleason score and tumor stage, data from the TCGA data set (queried from cBioPortal). LanCL1 expression is exon-normalized signal intensity. Mann–Whitney U -tests were performed to assess statistical significance for the comparisons between groups

Article Snippet: Fig. 2 Associations between LanCL1 expression and prostate cancer risk and severity. a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues.

Techniques: Expressing, Western Blot, Control, Quantitation Assay, MANN-WHITNEY

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a Representative immunohistochemistry of LanCL1 on benign prostatic epithelia (benign) and prostate cancer tissues (Gleason score 6 or 9). Student’s t-test was performed for statistical significance analysis. b Quantitation of protein expression of LanCL1 in benign or prostate cancer tissues. c Western blot analysis of LanCL1 expression in 15 pairs of nontumor tissues (P) and PCa tissues (T). Student’s t-test was performed for statistical significance analysis. N = 15. d Immunohistochemical staining for LanCL1 protein in the dorsolateral prostate of WT and TRAMP mice

Article Snippet: Fig. 2 Associations between LanCL1 expression and prostate cancer risk and severity. a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues.

Techniques: Immunohistochemistry, Quantitation Assay, Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a Stably transfected LNCaP cells and PC-3 cells interfered with LanCL1 were established. LanCL1 was knockdown by siRNA as indicated. The efficiency of knockdown were examined by western blotting. Tubulin served as the control. b Cell proliferation was measured at the indicated time points by MTS colorimetric assay (absorbance at 490 nm (OD490). Student’s t -test was performed for statistical significance analysis. N >3. * P <0.05, ** P <0.01, *** P <0.001 c Tumor growth in LanCL1 overexpressed PC-3 cells and control cells was investigated by xenograft tumor models. Student’s t-test was performed for statistical significance analysis. N = 7. ** P <0.01, *** P <0.001. d The cell cycle distribution was analyzed by fluorescent-activated cell scanning (FACS) in LanCL1 overexpressed LNCaP cells. e Column chart indicates the mean and SEM of the S-phase fraction for each cell. Student’s t-test was performed for statistical significance analysis. N = 5

Article Snippet: Fig. 2 Associations between LanCL1 expression and prostate cancer risk and severity. a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues.

Techniques: Stable Transfection, Transfection, Knockdown, Western Blot, Control, Colorimetric Assay

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a qRT–PCR shows no induction of LanCL1 mRNA in H 2 O 2 -treated (100 μM, 1 h) LNCaP cells. Student’s t -test was performed for statistical significance analysis. N = 3. b , c Western blots and quantification show that H 2 O 2 -treatment did not induce LanCL1 protein expression. Student’s t-test was performed for statistical significance analysis. N = 5. d – f Western blots and quantification show an increase in the level of 4-HNE in the H 2 O 2 -treated (100 μM, 1 h) LNCaP cells, but LanCL1 overexpression did not influence the level of 4-HNE. Western blots show that LanCL1 overexpression in LNCaP cells is not more resistant to H 2 O 2 -induced 4-HNE accumulation. Student’s t-test was performed for statistical significance analysis. N = 5. g ROS determination by using a fluorescent probe DCFH-DA in LNCaP and PC-3 cells untreated and treated with H 2 O 2 (100 μM, 1 h). Student’s t-test was performed for statistical significance analysis. N = 3

Article Snippet: Fig. 2 Associations between LanCL1 expression and prostate cancer risk and severity. a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues.

Techniques: Quantitative RT-PCR, Western Blot, Expressing, Over Expression

Journal: Cell Death & Disease

Article Title: LanCL1 protects prostate cancer cells from oxidative stress via suppression of JNK pathway

doi: 10.1038/s41419-017-0207-0

Figure Lengend Snippet: a , b Modulation of apoptosis related signaling proteins using antibody array assay. Total protein was extracted from LNCaP-NEO and LNCaP-LanCL1 cells. A slide-based antibody array was used for simultaneous detection of 19 signaling molecules involved in stress response and apoptosis using a PathScan Stress and Apoptosis Signaling Array kit. Each protein was arranged in duplicate. c The proteins with changes concentration included pP44/42 MAPK (ERK1/2) (Thr202/Try204), pAkt(S473), pHSP27(Ser82), pSmad2(Ser465), pP53(Ser15), pP38 MAPK(Thr180/Try182), pSAPK/JNK(Thr183/Try185) and Cleaved-Caspase-3(Asp175), which were indicated on the images, and quantified. d The concentrations of the protein in c were quantified. Student’s t -test was performed for statistical significance analysis. N = 3. Data are presented as mean ± SEM. * P <0.05, ** P <0.01, *** P <0.001. e Luciferase assay shows that overexpression of LanCL1 reduced AP-1 transcription. Student’s t-test was performed for statistical significance analysis. N = 3. *** P <0.001 f Western blotting indicated the protein level of pSAPK/JNK(Thr183/Try185) in overexpression LNCaP and PC-3 cells. Student’s t -test was performed for statistical significance analysis. N = 3. g Western blotting indicated the protein level of pSAPK/JNK(Thr183/Try185) in overexpression LNCaP cells after 100 μM H 2 O 2 for 2 h. Student’s t-test was performed for statistical significance analysis. N = 3. H. SP600125 partially rescued the cell death caused by ROS in LanCL1 knockdown LNCaP cells. Student’s t-test was performed for statistical significance analysis. N = 5

Article Snippet: Fig. 2 Associations between LanCL1 expression and prostate cancer risk and severity. a Data from The Human Protein Atlas portal show LanCL1 expression in different tissues.

Techniques: Ab Array, Concentration Assay, Luciferase, Over Expression, Western Blot, Knockdown

Journal: Developmental cell

Article Title: Developmental and Activity-Dependent Expression of LanCL1 Confers Antioxidant Activity Required for Neuronal Survival

doi: 10.1016/j.devcel.2014.06.011

Figure Lengend Snippet: (A) qRT-PCR shows induction of LanCL1 mRNA in bicuculline-treated (Bic, 5h) cortical neurons (DIV7). Error bars indicate SEM. **p= 0.0016. n=6.

Article Snippet: M.N.C., L.Z., Y.C., Y.Z. generated LanCL1 mouse models with the help of the Transgenic Facility of Johns Hopkins University.

Techniques: Quantitative RT-PCR

Journal: Developmental cell

Article Title: Developmental and Activity-Dependent Expression of LanCL1 Confers Antioxidant Activity Required for Neuronal Survival

doi: 10.1016/j.devcel.2014.06.011

Figure Lengend Snippet: (A and B) Photographs show that the genetic deletion of LanCL1 does not affect the body or brain size (LanCL1 −/−, ko, hereafter).

Article Snippet: M.N.C., L.Z., Y.C., Y.Z. generated LanCL1 mouse models with the help of the Transgenic Facility of Johns Hopkins University.

Techniques:

Journal: Developmental cell

Article Title: Developmental and Activity-Dependent Expression of LanCL1 Confers Antioxidant Activity Required for Neuronal Survival

doi: 10.1016/j.devcel.2014.06.011

Figure Lengend Snippet: (A) Representative images of ethidium fluorescence (Eth, red) show the accumulation of reactive oxygen species (ROS) in the cortex of 8-week LanCL1 ko brain. Bar:50μm.

Article Snippet: M.N.C., L.Z., Y.C., Y.Z. generated LanCL1 mouse models with the help of the Transgenic Facility of Johns Hopkins University.

Techniques: Fluorescence

Journal: Developmental cell

Article Title: Developmental and Activity-Dependent Expression of LanCL1 Confers Antioxidant Activity Required for Neuronal Survival

doi: 10.1016/j.devcel.2014.06.011

Figure Lengend Snippet: (A–D) Hoechst staining shows that deletion of LanCL1 increased neuronal death induced by H2O2 (12 hour treatment, DIV14, Ctrl: LanCL1 +/+, KO: LanCL1 −/−) (A, B), and LanCL1 transgene reduced neuronal death (12 hour treatment, DIV7, Ctrl: LanCL1 K/K, KI: LanCL1 K/K Nestin-Cre) (C, D). The data represent the mean ± SEM from four independent experiments, with a total number of 2,000 neurons analyzed for each group. Bar, 50μm. Error bars indicate SEM, *p= 0.0135, **p= 0.0069. n=4.

Article Snippet: M.N.C., L.Z., Y.C., Y.Z. generated LanCL1 mouse models with the help of the Transgenic Facility of Johns Hopkins University.

Techniques: Staining