Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Glucocorticoids suppress cystathionine gamma-lyase expression and H 2 S production in lipopolysaccharide-treated macrophages

doi: 10.1007/s00018-009-0250-9

Figure Lengend Snippet: CSE siRNA increased LPS-induced NO production and iNOS expression. RAW264.7 macrophages were separately transfected with pRNAT-U6.1/Neo-mCSE and pRNAT-U6.1/Neo-Control vectors. Stable CSE-knockdown RAW264.7 cells were selected and maintained in medium containing G418 and named “RAW-mCSE siRNA” (“RAW-control siRNA” as control). Western blot analysis (a) and real-time H2S production measurement (b) were used to determine CSE protein expression and H2S synthetic activity in CSE-knockdown RAW264.7 cells. After stimulation with LPS (1 μg/ml) for 24 h, NO production (c) and iNOS protein expression (d) were determined as described in “Materials and Methods.” Data were expressed as mean ± SEM for H2S production rates (n = 3) and NO production (n = 4), and mean percentage of control ± SEM for iNOS protein expression (n = 3). *P < 0.05; **P < 0.01 compared with “RAW-control siRNA”; ##P < 0.01 compared with “RAW-control siRNA” stimulated with LPS. Control siRNA: RAW-control siRNA; mCSE siRNA: RAW-mCSE siRNA

Article Snippet: The mouse CSE small interfering RNA (siRNA) plasmid was constructed using the pRNAT-U6.1/Neo vector (Jinsite Biotechnology) for knockdown of the CSE gene in vitro.

Techniques: Expressing, Transfection, Western Blot, Activity Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Glucocorticoids suppress cystathionine gamma-lyase expression and H 2 S production in lipopolysaccharide-treated macrophages

doi: 10.1007/s00018-009-0250-9

Figure Lengend Snippet: Effects of dexamethasone on CSE mRNA stability in macrophages stimulated with LPS. Primary (a) and RAW 264.7 (b) macrophages were stimulated with LPS (1 μg/ml) in the absence or presence of dexamethasone (10−7 mol/l) as indicated. Actinomycin D (1 μg/ml) was added to the cells after 6 h of incubation. Cells treated with vehicle were used as the maximum point. Cells were harvested for RNA extraction after the incubation time indicated after actinomycin D addition, and CSE and GAPDH mRNA were detected by quantitative real-time RT-PCR. Each data point was expressed as mean percentage of the maximum determined at time zero ± SEM (n = 4). Dex dexamethasone

Article Snippet: The mouse CSE small interfering RNA (siRNA) plasmid was constructed using the pRNAT-U6.1/Neo vector (Jinsite Biotechnology) for knockdown of the CSE gene in vitro.

Techniques: Incubation, RNA Extraction, Quantitative RT-PCR

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: LINC00511 was upregulated in LUAD. A The expression of LINC00511 was detected by qRT-PCR assay in Human LUAD cell line (A549, Calu-3, DV-90 and PC-9) and human bronchial epithelial cell line BEAS-2B. ** P < 0.001 compared with BEAS-2B. B The expression of LINC00511 was detected by qRT-PCR assay in cancer tissues ( n = 40) and normal tissues ( n = 40). C The subcellular localization assay for LINC00511. D The expression of LINC00511 was tested by qRT-PCR in A549 and PC9 cells transfected with si-NC and si-lnc for 48 h. ** P < 0.001 compared with si-NC. si-lnc, siRNA of LINC00511. si-NC, negative control of si-RNA

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Negative Control

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: Interference with LINC00511 inhibited the malignant proliferation of LUAD ells. A Cell viability was detected by CCK-8 assay in A549 and PC9 cells transfected with si-NC and si-lnc for 0, 24, 48 or 72 h. B Cell proliferation was determined by BrdU assay in A549 and PC9 cells transfected with si-NC and si-lnc for 48 h. C Bax and Bl-2 protein expression detected by western blot assay in A549 and PC9 cells transfected with si-NC and si-lnc for 48 h. D Migration capacity was determined by wound healing assay in A549 and PC9 cells transfected with si-NC and si-lnc for 48 h. (E) Cell invasion ability was detected by transwell invasion assay in A549 and PC9 cells transfected with si-NC and si-lnc for 48 h. ** P < 0.001 compared with si-NC. si-lnc, siRNA of LINC00511. si-NC, negative control of si-RNA

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: CCK-8 Assay, Transfection, BrdU Staining, Expressing, Western Blot, Migration, Wound Healing Assay, Transwell Invasion Assay, Negative Control

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: LINC00511 targeted and negatively regulated miR-195-5p. A The binding site of LINC00511 with miR-195-5p was confirmed according to bioinformatics analysis. B Luciferase reporter assay confirmed the molecular binding. ** P < 0.001 compared with mimic-NC. C RIP was conducted to measure the enrichment of LINC00511 in Ago2 immunoprecipitate and IgG-pellet. ** P < 0.001. D The expression of miR-195-5p was detected by qRT-PCR assay in cancer tissues ( n = 40) and normal tissues ( n = 40). ** P < 0.001. E pearson analysis was used to analyze the expression of LINC00511 and miR-195-5p in cancer tissues. F The expression of miR-195-5p was detected by qRT-PCR assay in A549, PC9 and BEAS-2B cells. ** P < 0.001 compared with BEAS-2B. G The LINC00511 and miR-195-5p expression were detected by qRT-PCR assay in A549 and PC9 cells transfected with si-lnc or inhibitor for 48 h. ** P < 0.001 compared with si-NC group. $$P < 0.001 compared with inhibitor-NC. ## P < 0.001 compared with si-lnc + inhibitor. si-lnc, siRNA of LINC00511. si-NC, negative control of siRNA. inhibitor, miR-195-5p inhibitor. inhibitor-NC, negative control of inhibitor. si-lnc + inhibitor, siRNA of LINC00511 + miR-195-5p inhibitor

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR, Transfection, Negative Control

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: LINC00511 negatively regulated miR-195-5p to affect the proliferation and apoptosis of LUAD cells. A Cell viability was detected by CCK-8 assay in A549 and PC9 cells transfected with si-lnc or inhibitor for 0, 24, 48 or 72 h. B Cell proliferation was determined by BrdU assay in A549 and PC9 cells transfected with si-lnc or inhibitor for 48 h. C . Bax and Bl-2 protein expression detected by western blot assay in A549 and PC9 cells transfected with si-lnc or inhibitor for 48 h. ** P < 0.001 compared with si-NC group. $ P < 0.05, $$ P < 0.001 compared with inhibitor-NC. # P < 0.05, ## P < 0.001 compared with si-lnc + inhibitor. si-lnc, siRNA of LINC00511. si-NC, negative control of siRNA. inhibitor, miR-195-5p inhibitor. inhibitor-NC, negative control of inhibitor. si-lnc + inhibitor, siRNA of LINC00511 + miR-195-5p inhibitor

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: CCK-8 Assay, Transfection, BrdU Staining, Expressing, Western Blot, Negative Control

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: LINC00511 negatively regulated miR-195-5p to affect the metastasis of LUAD cells. A Migration capacity was determined by wound healing assay in A549 and PC9 cells transfected with si-lnc or inhibitor for 48 h. B Cell invasion ability was detected by transwell invasion assay in A549 and PC9 cells transfected with si-lnc or inhibitor for 48 h. * P < 0.05, ** P < 0.001 compared with si-NC group. $$ P < 0.001 compared with inhibitor-NC. # P < 0.05, ## P < 0.001 compared with si-lnc + inhibitor. si-lnc, siRNA of LINC00511. si-NC, negative control of siRNA. inhibitor, miR-195-5p inhibitor. inhibitor-NC, negative control of inhibitor. si-lnc + inhibitor, siRNA of LINC00511 + miR-195-5p inhibitor

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: Migration, Wound Healing Assay, Transfection, Transwell Invasion Assay, Negative Control

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: GCNT3 was the target gene of miR-195-5p. A The schematic diagram presents the complementary binding sites within GCNT3 and miR-195-5p. B Luciferase reporter assay confirmed the molecular binding. ** P < 0.001 compared with WT + NC. # P < 0.05 compared with WT + mimic. C The expression of GCNT3 was detected by qRT-PCR assay in cancer tissues and normal tissues. ** P < 0.001. D Pearson analysis was used to analyze the expression of GCNT3 and miR-195-5p in cancer tissues. E The expression of GCNT3 was detected by qRT-PCR assay in A549, PC9 and BEAS-2B cells. ** P < 0.001 compared with BEAS-2B. F The GCNT3 mRNA expression were detected by qRT-PCR assay in A549 and PC9 cells transfected with si-lnc for 48 h. G The GCNT3 protein expression were detected by western blot assay in A549 and PC9 cells transfected with si-lnc for 48 h. H The GCNT3 protein expression were detected by western blot assay in A549 and PC9 cells transfected with si-GCNT3 or inhibitor for 48 h. ** P < 0.001 compared with si-NC group. $$ P < 0.001 compared with inhibitor-NC. ## P < 0.001 compared with si-GCNT3 + inhibitor. si-GCNT3, siRNA of GCNT3. si-NC, negative control of siRNA. inhibitor, miR-195-5p inhibitor. inhibitor-NC, negative control of inhibitor. si-GCNT3 + inhibitor, siRNA of GCNT3 + miR-195-5p inhibitor

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR, Transfection, Western Blot, Negative Control

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: GCNT3 silencing reversed the effect of miR-195-5p on the proliferation and apoptosis of LUAD cells. A Cell viability was detected by CCK-8 assay in A549 and PC9 cells transfected with si-GCNT3 or inhibitor for 0, 24, 48 or 72 h. B Cell proliferation was determined by BrdU assay in A549 and PC9 cells transfected with si-GCNT3 or inhibitor for 48 h. C Bax and Bl-2 protein expression detected by western blot assay in A549 and PC9 cells transfected with si-GCNT3 or inhibitor for 48 h. *P < 0.05, ** P < 0.001 compared with si-NC group. $ P < 0.05, $$ P < 0.001 compared with inhibitor-NC. # P < 0.05, ## P < 0.001 compared with si-GCNT3 + inhibitor. si-GCNT3, siRNA of GCNT3. si-NC, negative control of siRNA. inhibitor, miR-195-5p inhibitor. inhibitor-NC, negative control of inhibitor. si-GCNT3 + inhibitor, siRNA of GCNT3 + miR-195-5p inhibitor

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: CCK-8 Assay, Transfection, BrdU Staining, Expressing, Western Blot, Negative Control

Journal: BMC Cancer

Article Title: LINC00511 enhances LUAD malignancy by upregulating GCNT3 via miR-195-5p

doi: 10.1186/s12885-022-09459-7

Figure Lengend Snippet: GCNT3 silencing reversed the effect of miR-195-5p on the p metastasis of LUAD cells. A Migration capacity was determined by wound healing assay in A549 and PC9 cells transfected with si-GCNT3 or inhibitor for 48 h. B Cell invasion ability was detected by transwell invasion assay in A549 and PC9 cells transfected with si-GCNT3 or inhibitor for 48 h. ** P < 0.001 compared with si-NC group. $$ P < 0.001 compared with inhibitor-NC. ## P < 0.001 compared with si-GCNT3 + inhibitor. si-GCNT3, siRNA of GCNT3. si-NC, negative control of siRNA. inhibitor, miR-195-5p inhibitor. inhibitor-NC, negative control of inhibitor. si-GCNT3 + inhibitor, siRNA of GCNT3 + miR-195-5p inhibitor

Article Snippet: Specific siRNAs targeting LINC00511 (si-LNC), and GCNT3 (si-GCNT3), and control siRNA plasmid (si-NC) were synthesized by RiboBio (Guangzhou, China).

Techniques: Migration, Wound Healing Assay, Transfection, Transwell Invasion Assay, Negative Control