Journal: International journal of molecular sciences

Article Title: miR-212-5p Regulates PM 2.5 -Induced Apoptosis by Targeting LAMC2 and LAMA3.

doi: 10.3390/ijms26041761

Figure Lengend Snippet: Figure 3. miR-212-5p negatively regulates LAMC2 and LAMA3: (A) miRanda (v3.3a) and TargetScan (v5.0) projected miR-212-5p target genes displayed; (B) similarity of rno-miR-212-5p and has-miR- 212-5p genes shown; (C) schematic representation of LAMC2-3′ UTR and LAMA3-3′ UTR with miR- 212-5p target prediction and reporter plasmid construction, where red characters indicate binding sites and ”*” characters indicate mutation sites; (D) luciferase activity assessed in 293T cells after co- transfection of LAMC2-WT, LAMC2-MUT, LAMA3-WT, and LAMA3-MUT with miR-212-5p mimics or miR-mimics NC for 48 h; (E) LAMC2-mVenus-WT, LAMC2-mVenus-MUT, LAMA3-mVenus-WT, and LAMA3-mVenus-MUT co-transfected with miR-mimics NC or miR-212-5p in 293T cells (scale bar: 300 µm); (F) RT-PCR assay for miR-212-5p mimics and miR-212-5p inhibitors on LAMC2 and LAMA3 expression after transfection; (G) Western blotting used to identify alterations in the LAMC2 and LAMA3 proteins. The mean ± standard deviation (n = 3) is used to express the data. ns, no significance; *, p < 0.05; **, p < 0.01; and ***, p < 0.001 indicate significance.

Article Snippet: Next, 5% skimmed milk (Beyotime Biotechnology, Shanghai, China) was blocked for 2 h, and the membranes were incubated with specific primary antibodies overnight at 4 ◦C: BAD, BCL-2, caspase-3, LAMC2, p85, AKT, p-AKT, IκBα, β-action (1:1000 dilution, Proteintech, Rosemont, IL, USA), p-IκBα, p65, p-p65 (1:500 dilution, Proteintech, IL, USA), LAMA3, p-p85 (1:500 dilution, Abways, Shanghai, China), and GAPDH (1:7000 dilution, Abways, Shanghai, China).

Techniques: Plasmid Preparation, Binding Assay, Mutagenesis, Luciferase, Activity Assay, Cotransfection, Transfection, Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Standard Deviation

Journal: International journal of molecular sciences

Article Title: miR-212-5p Regulates PM 2.5 -Induced Apoptosis by Targeting LAMC2 and LAMA3.

doi: 10.3390/ijms26041761

Figure Lengend Snippet: Figure 4. PM2.5-induced changes in LAMC2 and LAMA3 expression: (A) LAMC2 and LAMA3 gene expression detected with RT-PCR after PM2.5 stimulation; (B) LAMC2 and LAMA3 protein changes detected with Western blotting; (C) silencing efficiency of LAMC2 and LAMA3 after transfection with siRNA−LAMC2 or siRNA−LAMA3 detected with RT-PCR; (D) LAMC2 and LAMA3 protein changes detected with Western blotting. The mean ± standard deviation (n = 3) is used to express the data. ns, no significance; *, p < 0.05; **, p < 0.01; and ***, p < 0.001 indicate significance.

Article Snippet: Next, 5% skimmed milk (Beyotime Biotechnology, Shanghai, China) was blocked for 2 h, and the membranes were incubated with specific primary antibodies overnight at 4 ◦C: BAD, BCL-2, caspase-3, LAMC2, p85, AKT, p-AKT, IκBα, β-action (1:1000 dilution, Proteintech, Rosemont, IL, USA), p-IκBα, p65, p-p65 (1:500 dilution, Proteintech, IL, USA), LAMA3, p-p85 (1:500 dilution, Abways, Shanghai, China), and GAPDH (1:7000 dilution, Abways, Shanghai, China).

Techniques: Expressing, Gene Expression, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection, Standard Deviation

Journal: International journal of molecular sciences

Article Title: miR-212-5p Regulates PM 2.5 -Induced Apoptosis by Targeting LAMC2 and LAMA3.

doi: 10.3390/ijms26041761

Figure Lengend Snippet: Figure 5. LAMC2 and LAMA3 inhibit apoptosis through the PI3K/AKT/NF-κB pathway: (A) PM2.5- stimulated (180 µg/mL for 24 h) cells transfected with siRNA−LAMC2 or siRNA−LAMA3, and p-IκBα/IκBα and p-p65/p65 protein changes; (B) p-AKT/AKT and p-p85/p85 protein changes; (C) BCL-2, BAD, and caspase-3 protein changes detected with Western blotting; (D) apoptosis rate detected with flow cytometry. The mean ± standard deviation (n = 3) is used to express the data. ns, no significance; *, p < 0.05; **, p < 0.01; and ***, p < 0.001 indicate significance.

Article Snippet: Next, 5% skimmed milk (Beyotime Biotechnology, Shanghai, China) was blocked for 2 h, and the membranes were incubated with specific primary antibodies overnight at 4 ◦C: BAD, BCL-2, caspase-3, LAMC2, p85, AKT, p-AKT, IκBα, β-action (1:1000 dilution, Proteintech, Rosemont, IL, USA), p-IκBα, p65, p-p65 (1:500 dilution, Proteintech, IL, USA), LAMA3, p-p85 (1:500 dilution, Abways, Shanghai, China), and GAPDH (1:7000 dilution, Abways, Shanghai, China).

Techniques: Transfection, Western Blot, Flow Cytometry, Standard Deviation

Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: Inactivation curves for M. avium subsp. paratuberculosis in acetate buffer (50 mM, pH 4) with various salt concentrations (0, 2, 4, and 6% [wt/vol]) at 20°C. (A) Dominic. (B) ATCC 19698.

Article Snippet: M. avium subsp. paratuberculosis strains were not viable beyond 56 days in all salt concentrations tested, but ATCC 19698 in the absence of salt was viable for up to 112 days at pH 4 (Fig. B). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 1 caption a7 Inactivation curves for M. avium subsp. paratuberculosis in acetate buffer (50 mM, pH 4) with various salt concentrations (0, 2, 4, and 6% [wt/vol]) at 20°C. (A) Dominic. (B) ATCC 19698. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 pH NaCl concn (%) D value (days) for strain: ATCC 19698 Dominic 4 0 16.1 a 9.2 2 7.9 9.7 4 7.8 9.9 6 8.7 11.3 5 0 27.2 a 19.7 2 17.4 19.6 4 19.3 19.7 6 17.5 11.8 a 6 0 32.5 31.9 2 38.2 32.0 4 35.2 24.0 6 39.1 33.9 Open in a separate window a Significantly different from D values for other individual NaCl concentrations at the same pH.

Techniques:

Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: D values for M. avium subsp. paratuberculosis strains tested at pH 4, 5, and 6 at 20°C

Article Snippet: M. avium subsp. paratuberculosis strains were not viable beyond 56 days in all salt concentrations tested, but ATCC 19698 in the absence of salt was viable for up to 112 days at pH 4 (Fig. B). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 1 caption a7 Inactivation curves for M. avium subsp. paratuberculosis in acetate buffer (50 mM, pH 4) with various salt concentrations (0, 2, 4, and 6% [wt/vol]) at 20°C. (A) Dominic. (B) ATCC 19698. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 pH NaCl concn (%) D value (days) for strain: ATCC 19698 Dominic 4 0 16.1 a 9.2 2 7.9 9.7 4 7.8 9.9 6 8.7 11.3 5 0 27.2 a 19.7 2 17.4 19.6 4 19.3 19.7 6 17.5 11.8 a 6 0 32.5 31.9 2 38.2 32.0 4 35.2 24.0 6 39.1 33.9 Open in a separate window a Significantly different from D values for other individual NaCl concentrations at the same pH.

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Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: D values for heat-treated M. avium subsp. paratuberculosis strains at pH 5, 2% (vol/vol) NaCl, and 20°C

Article Snippet: M. avium subsp. paratuberculosis strains were not viable beyond 56 days in all salt concentrations tested, but ATCC 19698 in the absence of salt was viable for up to 112 days at pH 4 (Fig. B). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG. 1 caption a7 Inactivation curves for M. avium subsp. paratuberculosis in acetate buffer (50 mM, pH 4) with various salt concentrations (0, 2, 4, and 6% [wt/vol]) at 20°C. (A) Dominic. (B) ATCC 19698. table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 pH NaCl concn (%) D value (days) for strain: ATCC 19698 Dominic 4 0 16.1 a 9.2 2 7.9 9.7 4 7.8 9.9 6 8.7 11.3 5 0 27.2 a 19.7 2 17.4 19.6 4 19.3 19.7 6 17.5 11.8 a 6 0 32.5 31.9 2 38.2 32.0 4 35.2 24.0 6 39.1 33.9 Open in a separate window a Significantly different from D values for other individual NaCl concentrations at the same pH.

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Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: Inactivation curves for M. avium subsp. paratuberculosis in acetate buffer (50 mM, pH 4) with various salt concentrations (0, 2, 4, and 6% [wt/vol]) at 20°C. (A) Dominic. (B) ATCC 19698.

Article Snippet: Thus, heat treatment in this buffer (at 62 or 71°C) resulted in D values for both strains that were lower than the D values for cells not exposed to heat. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Temp (°C) Heating time (s) Log 10 cell no. (mean ± SD) after heat treatment of strain: D value (days) for strain: ATCC 19698 a Dominic b ATCC 19698 Dominic 62 0 6.26 ± 0.07 4.60 ± 0.05 13.2 12.3 240 4.87 ± 0.21 2.48 ± 0.20 3.1 4.3 480 4.26 ± 0.04 0.17 ± 0.23 2.4 NA b 720 3.86 ± 0.09 0 1.3 NA 960 2.27 ± 1.01 0 NA NA 71 0 6.65 ± 0.02 4.67 ± 0.13 16.3 17.4 20 6.03 ± 0.16 3.94 ± 0.04 5.9 4.7 40 5.11 ± 0.12 2.49 ± 0.26 2.7 1.4 60 3.83 ± 0.49 0 0.8 NA Open in a separate window a The D 62°C and D 71°C were 267.4 and 21.3 s, respectively. b The D 62°C and D 71°C were 108.4 and 13.5 s, respectively. c NA, D values were not available, since the number of M. avium subsp. paratuberculosis cells was low after heat treatment and the cells were completely inactivated at the first sampling time.

Techniques:

Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: D values for M. avium subsp. paratuberculosis strains tested at pH 4, 5, and 6 at 20°C

Article Snippet: Thus, heat treatment in this buffer (at 62 or 71°C) resulted in D values for both strains that were lower than the D values for cells not exposed to heat. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Temp (°C) Heating time (s) Log 10 cell no. (mean ± SD) after heat treatment of strain: D value (days) for strain: ATCC 19698 a Dominic b ATCC 19698 Dominic 62 0 6.26 ± 0.07 4.60 ± 0.05 13.2 12.3 240 4.87 ± 0.21 2.48 ± 0.20 3.1 4.3 480 4.26 ± 0.04 0.17 ± 0.23 2.4 NA b 720 3.86 ± 0.09 0 1.3 NA 960 2.27 ± 1.01 0 NA NA 71 0 6.65 ± 0.02 4.67 ± 0.13 16.3 17.4 20 6.03 ± 0.16 3.94 ± 0.04 5.9 4.7 40 5.11 ± 0.12 2.49 ± 0.26 2.7 1.4 60 3.83 ± 0.49 0 0.8 NA Open in a separate window a The D 62°C and D 71°C were 267.4 and 21.3 s, respectively. b The D 62°C and D 71°C were 108.4 and 13.5 s, respectively. c NA, D values were not available, since the number of M. avium subsp. paratuberculosis cells was low after heat treatment and the cells were completely inactivated at the first sampling time.

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Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: D values for heat-treated M. avium subsp. paratuberculosis strains at pH 5, 2% (vol/vol) NaCl, and 20°C

Article Snippet: Thus, heat treatment in this buffer (at 62 or 71°C) resulted in D values for both strains that were lower than the D values for cells not exposed to heat. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Temp (°C) Heating time (s) Log 10 cell no. (mean ± SD) after heat treatment of strain: D value (days) for strain: ATCC 19698 a Dominic b ATCC 19698 Dominic 62 0 6.26 ± 0.07 4.60 ± 0.05 13.2 12.3 240 4.87 ± 0.21 2.48 ± 0.20 3.1 4.3 480 4.26 ± 0.04 0.17 ± 0.23 2.4 NA b 720 3.86 ± 0.09 0 1.3 NA 960 2.27 ± 1.01 0 NA NA 71 0 6.65 ± 0.02 4.67 ± 0.13 16.3 17.4 20 6.03 ± 0.16 3.94 ± 0.04 5.9 4.7 40 5.11 ± 0.12 2.49 ± 0.26 2.7 1.4 60 3.83 ± 0.49 0 0.8 NA Open in a separate window a The D 62°C and D 71°C were 267.4 and 21.3 s, respectively. b The D 62°C and D 71°C were 108.4 and 13.5 s, respectively. c NA, D values were not available, since the number of M. avium subsp. paratuberculosis cells was low after heat treatment and the cells were completely inactivated at the first sampling time.

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Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: Inactivation curves for nonheated and heat-treated (at 62°C for 240 s) M. avium subsp. paratuberculosis strain ATCC 19698 in the acetate buffer (pH 5, 2% [wt/vol] NaCl) at 20°C.

Article Snippet: Thus, heat treatment in this buffer (at 62 or 71°C) resulted in D values for both strains that were lower than the D values for cells not exposed to heat. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Temp (°C) Heating time (s) Log 10 cell no. (mean ± SD) after heat treatment of strain: D value (days) for strain: ATCC 19698 a Dominic b ATCC 19698 Dominic 62 0 6.26 ± 0.07 4.60 ± 0.05 13.2 12.3 240 4.87 ± 0.21 2.48 ± 0.20 3.1 4.3 480 4.26 ± 0.04 0.17 ± 0.23 2.4 NA b 720 3.86 ± 0.09 0 1.3 NA 960 2.27 ± 1.01 0 NA NA 71 0 6.65 ± 0.02 4.67 ± 0.13 16.3 17.4 20 6.03 ± 0.16 3.94 ± 0.04 5.9 4.7 40 5.11 ± 0.12 2.49 ± 0.26 2.7 1.4 60 3.83 ± 0.49 0 0.8 NA Open in a separate window a The D 62°C and D 71°C were 267.4 and 21.3 s, respectively. b The D 62°C and D 71°C were 108.4 and 13.5 s, respectively. c NA, D values were not available, since the number of M. avium subsp. paratuberculosis cells was low after heat treatment and the cells were completely inactivated at the first sampling time.

Techniques:

Journal:

Article Title: Effect of Three Factors in Cheese Production (pH, Salt, and Heat) on Mycobacterium avium subsp. paratuberculosis Viability

doi:

Figure Lengend Snippet: Inactivation curves for nonheated and heat-treated (at 62°C for 240 s) M. avium subsp. paratuberculosis strain ATCC 19698 in the soft white cheese at 4°C.

Article Snippet: Thus, heat treatment in this buffer (at 62 or 71°C) resulted in D values for both strains that were lower than the D values for cells not exposed to heat. table ft1 table-wrap mode="anchored" t5 TABLE 2 caption a7 Temp (°C) Heating time (s) Log 10 cell no. (mean ± SD) after heat treatment of strain: D value (days) for strain: ATCC 19698 a Dominic b ATCC 19698 Dominic 62 0 6.26 ± 0.07 4.60 ± 0.05 13.2 12.3 240 4.87 ± 0.21 2.48 ± 0.20 3.1 4.3 480 4.26 ± 0.04 0.17 ± 0.23 2.4 NA b 720 3.86 ± 0.09 0 1.3 NA 960 2.27 ± 1.01 0 NA NA 71 0 6.65 ± 0.02 4.67 ± 0.13 16.3 17.4 20 6.03 ± 0.16 3.94 ± 0.04 5.9 4.7 40 5.11 ± 0.12 2.49 ± 0.26 2.7 1.4 60 3.83 ± 0.49 0 0.8 NA Open in a separate window a The D 62°C and D 71°C were 267.4 and 21.3 s, respectively. b The D 62°C and D 71°C were 108.4 and 13.5 s, respectively. c NA, D values were not available, since the number of M. avium subsp. paratuberculosis cells was low after heat treatment and the cells were completely inactivated at the first sampling time.

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