rabbit anti-lc3 polyclonal antibody: pm036 Search Results


lc3  (MBL International)
90
MBL International lc3
Lc3, supplied by MBL International, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 1 article reviews
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anti-lc3  (MBL Life science)
90
MBL Life science anti-lc3
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Anti Lc3, supplied by MBL Life science, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-lc3/product/MBL Life science
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anti-lc3 wb  (Millipore)
90
Millipore anti-lc3 wb
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Anti Lc3 Wb, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sc 18822 rrid ab 626858 rabbit monoclonal anti tfam  (Santa Cruz Biotechnology)
96
Santa Cruz Biotechnology sc 18822 rrid ab 626858 rabbit monoclonal anti tfam
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Sc 18822 Rrid Ab 626858 Rabbit Monoclonal Anti Tfam, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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sc 18822 rrid ab 626858 rabbit monoclonal anti tfam - by Bioz Stars, 2026-03
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anti-lc3  (MBL International)
90
MBL International anti-lc3
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Anti Lc3, supplied by MBL International, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-lc3/product/MBL International
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rabbit anti-lc3 pm036  (Nordic BioSite)
90
Nordic BioSite rabbit anti-lc3 pm036
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Rabbit Anti Lc3 Pm036, supplied by Nordic BioSite, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rabbit anti-lc3 pm036 - by Bioz Stars, 2026-03
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anti lc3  (Proteintech)
93
Proteintech anti lc3
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Anti Lc3, supplied by Proteintech, 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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anti lc3 ii  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc anti lc3 ii
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Anti Lc3 Ii, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti lc3  (Cell Signaling Technology Inc)
97
Cell Signaling Technology Inc anti lc3
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Anti Lc3, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti lc3/product/Cell Signaling Technology Inc
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anti-lc3 4e12  (MBL International)
90
MBL International anti-lc3 4e12
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Anti Lc3 4e12, supplied by MBL International, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti-lc3 4e12/product/MBL International
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antibodies against lamin b1 pm064  (MBL Life science)
90
MBL Life science antibodies against lamin b1 pm064
Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, <t>LC3,</t> and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).
Antibodies Against Lamin B1 Pm064, supplied by MBL Life science, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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anti lc3 polyclonal antibody pm036  (Cell Signaling Technology Inc)
99
Cell Signaling Technology Inc anti lc3 polyclonal antibody pm036
Vitamin B12 induced autophagy in RIN-m5F cells. ( A ) RIN-m5F cells were subjected to varying concentrations of vitamin B12, and cytotoxicity levels were assessed utilizing a CCK-8 kit assay. ( B ) The cells were treated with vitamin B12 concentrations of 2, 4, and 8 μM for 24 h and stained utilizing <t>an</t> <t>anti-LC3</t> antibody (scale = 5 μm). ( C ) The cells were cultured as described in section ( B ), and the mathematical statistical methods were conducted to calculate the number of autophagosomes in each cell. At least 30 cells were counted for statistical analysis utilizing the t -test. ( D , F , H ) The impact of vitamin B12 at 2, 4, and 8 μM concentrations on the expression of autophagy-related proteins (anti-LC3, anti-p62, anti-GAPDH, anti-S6K, and an-ti-p-S6K) in cells were carried out using Western blotting. ( E , G , I ) The LC3-II/LC3-I, p62/GAPDH, and p-S6K/S6K ratios in ( D , F , H ), respectively, were analyzed by integrated optical density (IOD) using Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01 reveals significant differences in comparison to the control group.
Anti Lc3 Polyclonal Antibody Pm036, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/anti lc3 polyclonal antibody pm036/product/Cell Signaling Technology Inc
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Image Search Results


Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, LC3, and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).

Journal: bioRxiv

Article Title: Temporal inhibition of electron transport chain attenuates stress-induced cellular senescence by prolonged disturbance of proteostasis in human fibroblasts

doi: 10.1101/2022.11.07.515395

Figure Lengend Snippet: Antimycin A suppresses accumulation of intracellular protein aggregates induced by prolonged inhibition of proteostasis (A) The fluorescence intensity for intracellular protein aggregates was normalized to the number of nuclei. Relative values are shown as the mean (SD) from the measurements of the four different images. The asterisk (*) indicates p < 0.05 by a Mann-Whitney U test for MG132-treated (-) vs. AA co-treated (+) cells. (B) Representative images of accumulated protein aggregates in controls (39 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with ProteoStat Aggresome Detection Reagent (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (C) The fluorescence intensity for acidic lysosome was normalized to the number of nuclei. ns, not significant. (D) Representative images of acidic lysosomes in controls (41 PDL), MG132-treated, and AA co-treated cells on day 3. Cells were stained with AcidiFluor ORANGE (red) and Hoechst 33342 (blue). Scale bar = 50 µm. (E) Immunoblot analyses of HSP70, LC3, and p62/SQSTM1 for controls, MG132-treated (-), and AA co-treated (+) cells. Intensities of immunoblot bands of LC3-II (F) and HSP70 (G) were normalized to those of LC3-I or GAPDH, respectively. Relative values are shown as the mean (SD) ( n = 4).

Article Snippet: anti-S6 (C-8, sc-74459, Santa Cruz Biotechnology, mouse monoclonal) anti-p-S6 (50.Ser 235/236, sc-293144, Santa Cruz Biotechnology, mouse monoclonal) anti-NRF1 (G-5, sc-515360, Santa Cruz Biotechnology, mouse monoclonal) anti-NRF2 (M200-3, MBL Life Science, mouse monoclonal) anti-LC3 (PM036, MBL Life Science, rabbit polyclonal) anti-GAPDH (G8795, Sigma-aldrich, mouse monoclonal) anti-AKT (#9272S, Cell Signaling Technology, rabbit polyclonal) anti-p-AKT (S473) (#9271S, Cell Signaling Technology, rabbit polyclonal) anti-SQSTM1 (p62) (D-3, sc-28359, Santa Cruz Biotechnology, mouse monoclonal) anti-HSP70 (C92F3A-5, sc-66048, Santa Cruz Biotechnology, mouse monoclonal) anti-NIX (H-8, sc-166332, Santa Cruz Biotechnology, mouse monoclonal) anti-BNIP-3 (ANa40, sc-56167, Santa Cruz Biotechnology, mouse monoclonal) anti-Parkin (PRK8, sc-32282, Santa Cruz Biotechnology, mouse monoclonal) anti-TFAM (C-9, sc-376672, Santa Cruz Biotechnology, mouse monoclonal) anti-DRP1 (C-5, sc-271583, Santa Cruz Biotechnology, mouse monoclonal)

Techniques: Inhibition, Fluorescence, MANN-WHITNEY, Staining, Western Blot

Vitamin B12 induced autophagy in RIN-m5F cells. ( A ) RIN-m5F cells were subjected to varying concentrations of vitamin B12, and cytotoxicity levels were assessed utilizing a CCK-8 kit assay. ( B ) The cells were treated with vitamin B12 concentrations of 2, 4, and 8 μM for 24 h and stained utilizing an anti-LC3 antibody (scale = 5 μm). ( C ) The cells were cultured as described in section ( B ), and the mathematical statistical methods were conducted to calculate the number of autophagosomes in each cell. At least 30 cells were counted for statistical analysis utilizing the t -test. ( D , F , H ) The impact of vitamin B12 at 2, 4, and 8 μM concentrations on the expression of autophagy-related proteins (anti-LC3, anti-p62, anti-GAPDH, anti-S6K, and an-ti-p-S6K) in cells were carried out using Western blotting. ( E , G , I ) The LC3-II/LC3-I, p62/GAPDH, and p-S6K/S6K ratios in ( D , F , H ), respectively, were analyzed by integrated optical density (IOD) using Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01 reveals significant differences in comparison to the control group.

Journal: International Journal of Molecular Sciences

Article Title: Vitamin B12-Induced Autophagy Alleviates High Glucose-Mediated Apoptosis of Islet β Cells

doi: 10.3390/ijms242015217

Figure Lengend Snippet: Vitamin B12 induced autophagy in RIN-m5F cells. ( A ) RIN-m5F cells were subjected to varying concentrations of vitamin B12, and cytotoxicity levels were assessed utilizing a CCK-8 kit assay. ( B ) The cells were treated with vitamin B12 concentrations of 2, 4, and 8 μM for 24 h and stained utilizing an anti-LC3 antibody (scale = 5 μm). ( C ) The cells were cultured as described in section ( B ), and the mathematical statistical methods were conducted to calculate the number of autophagosomes in each cell. At least 30 cells were counted for statistical analysis utilizing the t -test. ( D , F , H ) The impact of vitamin B12 at 2, 4, and 8 μM concentrations on the expression of autophagy-related proteins (anti-LC3, anti-p62, anti-GAPDH, anti-S6K, and an-ti-p-S6K) in cells were carried out using Western blotting. ( E , G , I ) The LC3-II/LC3-I, p62/GAPDH, and p-S6K/S6K ratios in ( D , F , H ), respectively, were analyzed by integrated optical density (IOD) using Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01 reveals significant differences in comparison to the control group.

Article Snippet: The following antibodies were purchased from various sources: anti-LC3 polyclonal antibody (PM036), anti-LC3 monoclonal antibody (M186-3), and anti-p62 antibody (PM045) from Medical Biological Laboratory; anti-p70S6K antibody (2708) and anti-phosphorylated p70S6K antibody (9206) from Cell Signaling Technology (Beverly, MA, USA); and anti-3-phosphoglyceraldehyde dehydrogenase (GAPDH) anti-body (ZB002) from YTHX Biotechnology Co., Ltd. (Beijing, China).

Techniques: CCK-8 Assay, Staining, Cell Culture, Expressing, Western Blot, Software, Standard Deviation, Comparison, Control

The influence of high glucose on autophagy in RIN-m5F cells. ( A ) RIN-m5F cells were exposed to the indicated concentrations of glucose, and cytotoxicity was assessed utilizing a CCK-8 kit assay. ( B ) The cells were cultured under conditions of 35, 45, and 55 mM glucose for 36 h and then stained with anti-LC3 antibody (scale = 5 μm). ( C ) The cells were treated according to section ( A ), mathematical statistical methods were conducted to calculate the number of autophagosomes per cell, and at least 30 cells were counted by statistical analysis based on a t -test. ( D ) The cells were treated according section ( A ), and Western blotting analysis was conducted to investigate the expression levels of autophagy-related proteins with specific antibodies including anti-LC3, anti-p62, anti-GAPDH, anti-S6K, and anti-p-S6K. ( E , F ) Subsequent to the procedure described in section ( D ), the ratios of p62/GAPDH and p-S6K/S6K were assessed through IOD measurements utilizing Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01 reveals significant differences in comparison to the control group.

Journal: International Journal of Molecular Sciences

Article Title: Vitamin B12-Induced Autophagy Alleviates High Glucose-Mediated Apoptosis of Islet β Cells

doi: 10.3390/ijms242015217

Figure Lengend Snippet: The influence of high glucose on autophagy in RIN-m5F cells. ( A ) RIN-m5F cells were exposed to the indicated concentrations of glucose, and cytotoxicity was assessed utilizing a CCK-8 kit assay. ( B ) The cells were cultured under conditions of 35, 45, and 55 mM glucose for 36 h and then stained with anti-LC3 antibody (scale = 5 μm). ( C ) The cells were treated according to section ( A ), mathematical statistical methods were conducted to calculate the number of autophagosomes per cell, and at least 30 cells were counted by statistical analysis based on a t -test. ( D ) The cells were treated according section ( A ), and Western blotting analysis was conducted to investigate the expression levels of autophagy-related proteins with specific antibodies including anti-LC3, anti-p62, anti-GAPDH, anti-S6K, and anti-p-S6K. ( E , F ) Subsequent to the procedure described in section ( D ), the ratios of p62/GAPDH and p-S6K/S6K were assessed through IOD measurements utilizing Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01 reveals significant differences in comparison to the control group.

Article Snippet: The following antibodies were purchased from various sources: anti-LC3 polyclonal antibody (PM036), anti-LC3 monoclonal antibody (M186-3), and anti-p62 antibody (PM045) from Medical Biological Laboratory; anti-p70S6K antibody (2708) and anti-phosphorylated p70S6K antibody (9206) from Cell Signaling Technology (Beverly, MA, USA); and anti-3-phosphoglyceraldehyde dehydrogenase (GAPDH) anti-body (ZB002) from YTHX Biotechnology Co., Ltd. (Beijing, China).

Techniques: CCK-8 Assay, Cell Culture, Staining, Western Blot, Expressing, Software, Standard Deviation, Comparison, Control

Vitamin B12 induced autophagy under conditions of high glucose stress. ( A ) RIN-m5F cells were treated with varying concentrations of vitamin B12 (2, 4, and 8 μM) in conjunction with 45 mM glucose for 36 h, and the number of autophagosomes was determined through immunofluorescence analysis (scale = 5 μm). ( B ) The cells were treated according to section ( A ), and mathematical statistical methods were conducted to calculate the number of autophagosomes in each cell. At least 30 cells were counted by t test analysis. ( C , E , G ) The cells were treated according to section ( A ), and Western blotting was conducted to investigate the expression levels of proteins with specific anti-LC3, anti-p62, anti-GAPDH, anti-S6K, and anti-p-S6K antibodies. ( D , F , H ) The ratio of LC3-II/LC3-I, p62/GAPDH and p-S6K/S6K ratios in ( C , E , G ), respectively, were assessed through IOD measurements utilizing Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01, * p < 0.05 reveals significant differences in comparison to the control group.

Journal: International Journal of Molecular Sciences

Article Title: Vitamin B12-Induced Autophagy Alleviates High Glucose-Mediated Apoptosis of Islet β Cells

doi: 10.3390/ijms242015217

Figure Lengend Snippet: Vitamin B12 induced autophagy under conditions of high glucose stress. ( A ) RIN-m5F cells were treated with varying concentrations of vitamin B12 (2, 4, and 8 μM) in conjunction with 45 mM glucose for 36 h, and the number of autophagosomes was determined through immunofluorescence analysis (scale = 5 μm). ( B ) The cells were treated according to section ( A ), and mathematical statistical methods were conducted to calculate the number of autophagosomes in each cell. At least 30 cells were counted by t test analysis. ( C , E , G ) The cells were treated according to section ( A ), and Western blotting was conducted to investigate the expression levels of proteins with specific anti-LC3, anti-p62, anti-GAPDH, anti-S6K, and anti-p-S6K antibodies. ( D , F , H ) The ratio of LC3-II/LC3-I, p62/GAPDH and p-S6K/S6K ratios in ( C , E , G ), respectively, were assessed through IOD measurements utilizing Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01, * p < 0.05 reveals significant differences in comparison to the control group.

Article Snippet: The following antibodies were purchased from various sources: anti-LC3 polyclonal antibody (PM036), anti-LC3 monoclonal antibody (M186-3), and anti-p62 antibody (PM045) from Medical Biological Laboratory; anti-p70S6K antibody (2708) and anti-phosphorylated p70S6K antibody (9206) from Cell Signaling Technology (Beverly, MA, USA); and anti-3-phosphoglyceraldehyde dehydrogenase (GAPDH) anti-body (ZB002) from YTHX Biotechnology Co., Ltd. (Beijing, China).

Techniques: Immunofluorescence, Western Blot, Expressing, Software, Standard Deviation, Comparison, Control

3-MA suppressed vitamin B12-induced autophagy under the condition of high glucose stress. ( A ) RIN-m5F cells were cultivated under various conditions: either in the presence or absence of 45 mM glucose, supplemented with 10 mM 3-MA, a combination of 45 mM glucose and 10 mM 3-MA, 2 μM vitamin B12, 45 mM glucose plus 2 μM vitamin B12, or a concoction of 20 mM glucose, 2 μM vitamin B12, and 10 mM 3-MA, cultured for 36 h and stained with anti-LC3 anti-body (scale bar = 5 μm). ( B ) The cells were treated according to section ( A ), and mathematical statistical methods were conducted to calculate the number of autophagosomes in each cell. At least 30 cells were analyzed. ( C ) The cells were treated according to section ( A ), and Western blotting was performed to investigate the expression levels of proteins with specific antibodies against LC3 and GAPDH. ( D ) The LC3-II/LC3-I ratio in ( C ) was analyzed by IOD analysis utilizing Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01, * p < 0.05 reveals significant differences in comparison to the control group. ## p < 0.01 reveals significant differences in comparison to the group treated with vitamin B12 combined with high glucose through one-way ANOVA.

Journal: International Journal of Molecular Sciences

Article Title: Vitamin B12-Induced Autophagy Alleviates High Glucose-Mediated Apoptosis of Islet β Cells

doi: 10.3390/ijms242015217

Figure Lengend Snippet: 3-MA suppressed vitamin B12-induced autophagy under the condition of high glucose stress. ( A ) RIN-m5F cells were cultivated under various conditions: either in the presence or absence of 45 mM glucose, supplemented with 10 mM 3-MA, a combination of 45 mM glucose and 10 mM 3-MA, 2 μM vitamin B12, 45 mM glucose plus 2 μM vitamin B12, or a concoction of 20 mM glucose, 2 μM vitamin B12, and 10 mM 3-MA, cultured for 36 h and stained with anti-LC3 anti-body (scale bar = 5 μm). ( B ) The cells were treated according to section ( A ), and mathematical statistical methods were conducted to calculate the number of autophagosomes in each cell. At least 30 cells were analyzed. ( C ) The cells were treated according to section ( A ), and Western blotting was performed to investigate the expression levels of proteins with specific antibodies against LC3 and GAPDH. ( D ) The LC3-II/LC3-I ratio in ( C ) was analyzed by IOD analysis utilizing Image-Pro Plus 6.0 software. All experiments were conducted a minimum of three times. The error bars represent the standard deviation, ** p < 0.01, * p < 0.05 reveals significant differences in comparison to the control group. ## p < 0.01 reveals significant differences in comparison to the group treated with vitamin B12 combined with high glucose through one-way ANOVA.

Article Snippet: The following antibodies were purchased from various sources: anti-LC3 polyclonal antibody (PM036), anti-LC3 monoclonal antibody (M186-3), and anti-p62 antibody (PM045) from Medical Biological Laboratory; anti-p70S6K antibody (2708) and anti-phosphorylated p70S6K antibody (9206) from Cell Signaling Technology (Beverly, MA, USA); and anti-3-phosphoglyceraldehyde dehydrogenase (GAPDH) anti-body (ZB002) from YTHX Biotechnology Co., Ltd. (Beijing, China).

Techniques: Cell Culture, Staining, Western Blot, Expressing, Software, Standard Deviation, Comparison, Control