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rabbit polyclonal antibody against lin28a  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc rabbit polyclonal antibody against lin28a
    (A) An experimental timeline that describes injury, Lissamine-tagged Morpholino (MO) delivery, electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a decrease in the number of BrdU + retinal progenitors with yy1a MO and yy1b MO alone and in combination, compared to control MO injected retina at 4dpi (B), which is quantified (C). (D) Western Blot analyses of various RAGs in retinal extracts prepared from retinae electroporated with MOs against yy1a and yy1b at 2dpi. (E) The qPCR analyses of let-7a mi-RNA in the yy1a and yy1b knockdown condition at 2dpi; *p < 0.05; n=6 biological replicates. (F and G) The schematic showing Yy1-binding sites (BS) on ascl1a promoter (F) and the retinal ChIP assays confirm the physical binding of Yy1 on ascl1a promoter, in 2dpi retina (G). N.S marks the negative control. (H and I) Luciferase assay done in zebrafish embryos at 24hpf, injected with ascl1a: GFP-luciferase construct shows upregulation in the ascl1a promoter activity in yy1a and yy1b overexpression (H), while downregulation in their combined knockdown (I). ( J and K) The schematic showing Yy1-binding sites (BS) on <t>lin28a</t> promoter (J) and the retinal ChIP assays confirm the physical binding of Yy1 on lin28a promoter, in 2dpi retina (K). (L and M) Luciferase assay done in zebrafish embryos at 24hpf, injected with lin28a: GFP-luciferase construct shows upregulation in the lin28a promoter activity in yy1a and yy1b overexpression (L), while opposite in their combined knockdown (M). (N and O) The schematic showing Yy1-binding sites (BS) on yy1a intron (N) and the retinal ChIP assays confirm the loss of Yy1 binding onto its intron, in 2dpi retina (O). (P) The qPCR analysis shows the downregulation of yy1a and yy1b in their own combined knockdown. (Q and R) The schematic showing Yy1-binding sites (BS) on zic2b promoter (Q) and the retinal ChIP assays confirm the physical binding of Yy1 on zic2b promoter, in 2dpi retina (R). (S) Luciferase assay done in zebrafish embryos at 24hpf, injected with zic2b: GFP-luciferase construct shows upregulation in the zic2b promoter activity in yy1a and yy1b knockdown. (T and U) The schematic showing Yy1-binding sites (BS) on oct4 promoter (T) and the retinal ChIP assays confirm the physical binding of Yy1 on oct4 promoter, in 2dpi retina (U). (V) Luciferase assay done in zebrafish embryos at 24hpf, injected with oct4: GFP-luciferase construct shows upregulation in the oct4 promoter activity in yy1a and yy1b knockdown. Error bars represent SD. *p < 0.001 in (H, L, M, P, S, V); *p < 0.05 (F). n = 6 biological replicates (C, E and P).
    Rabbit Polyclonal Antibody Against Lin28a, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit polyclonal antibody against lin28a/product/Cell Signaling Technology Inc
    Average 95 stars, based on 125 article reviews
    rabbit polyclonal antibody against lin28a - by Bioz Stars, 2026-02
    95/100 stars

    Images

    1) Product Images from "YY1 protein is essential for the promotion of Muller glia reprogramming and retina regeneration"

    Article Title: YY1 protein is essential for the promotion of Muller glia reprogramming and retina regeneration

    Journal: bioRxiv

    doi: 10.1101/2025.01.31.635905

    (A) An experimental timeline that describes injury, Lissamine-tagged Morpholino (MO) delivery, electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a decrease in the number of BrdU + retinal progenitors with yy1a MO and yy1b MO alone and in combination, compared to control MO injected retina at 4dpi (B), which is quantified (C). (D) Western Blot analyses of various RAGs in retinal extracts prepared from retinae electroporated with MOs against yy1a and yy1b at 2dpi. (E) The qPCR analyses of let-7a mi-RNA in the yy1a and yy1b knockdown condition at 2dpi; *p < 0.05; n=6 biological replicates. (F and G) The schematic showing Yy1-binding sites (BS) on ascl1a promoter (F) and the retinal ChIP assays confirm the physical binding of Yy1 on ascl1a promoter, in 2dpi retina (G). N.S marks the negative control. (H and I) Luciferase assay done in zebrafish embryos at 24hpf, injected with ascl1a: GFP-luciferase construct shows upregulation in the ascl1a promoter activity in yy1a and yy1b overexpression (H), while downregulation in their combined knockdown (I). ( J and K) The schematic showing Yy1-binding sites (BS) on lin28a promoter (J) and the retinal ChIP assays confirm the physical binding of Yy1 on lin28a promoter, in 2dpi retina (K). (L and M) Luciferase assay done in zebrafish embryos at 24hpf, injected with lin28a: GFP-luciferase construct shows upregulation in the lin28a promoter activity in yy1a and yy1b overexpression (L), while opposite in their combined knockdown (M). (N and O) The schematic showing Yy1-binding sites (BS) on yy1a intron (N) and the retinal ChIP assays confirm the loss of Yy1 binding onto its intron, in 2dpi retina (O). (P) The qPCR analysis shows the downregulation of yy1a and yy1b in their own combined knockdown. (Q and R) The schematic showing Yy1-binding sites (BS) on zic2b promoter (Q) and the retinal ChIP assays confirm the physical binding of Yy1 on zic2b promoter, in 2dpi retina (R). (S) Luciferase assay done in zebrafish embryos at 24hpf, injected with zic2b: GFP-luciferase construct shows upregulation in the zic2b promoter activity in yy1a and yy1b knockdown. (T and U) The schematic showing Yy1-binding sites (BS) on oct4 promoter (T) and the retinal ChIP assays confirm the physical binding of Yy1 on oct4 promoter, in 2dpi retina (U). (V) Luciferase assay done in zebrafish embryos at 24hpf, injected with oct4: GFP-luciferase construct shows upregulation in the oct4 promoter activity in yy1a and yy1b knockdown. Error bars represent SD. *p < 0.001 in (H, L, M, P, S, V); *p < 0.05 (F). n = 6 biological replicates (C, E and P).
    Figure Legend Snippet: (A) An experimental timeline that describes injury, Lissamine-tagged Morpholino (MO) delivery, electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a decrease in the number of BrdU + retinal progenitors with yy1a MO and yy1b MO alone and in combination, compared to control MO injected retina at 4dpi (B), which is quantified (C). (D) Western Blot analyses of various RAGs in retinal extracts prepared from retinae electroporated with MOs against yy1a and yy1b at 2dpi. (E) The qPCR analyses of let-7a mi-RNA in the yy1a and yy1b knockdown condition at 2dpi; *p < 0.05; n=6 biological replicates. (F and G) The schematic showing Yy1-binding sites (BS) on ascl1a promoter (F) and the retinal ChIP assays confirm the physical binding of Yy1 on ascl1a promoter, in 2dpi retina (G). N.S marks the negative control. (H and I) Luciferase assay done in zebrafish embryos at 24hpf, injected with ascl1a: GFP-luciferase construct shows upregulation in the ascl1a promoter activity in yy1a and yy1b overexpression (H), while downregulation in their combined knockdown (I). ( J and K) The schematic showing Yy1-binding sites (BS) on lin28a promoter (J) and the retinal ChIP assays confirm the physical binding of Yy1 on lin28a promoter, in 2dpi retina (K). (L and M) Luciferase assay done in zebrafish embryos at 24hpf, injected with lin28a: GFP-luciferase construct shows upregulation in the lin28a promoter activity in yy1a and yy1b overexpression (L), while opposite in their combined knockdown (M). (N and O) The schematic showing Yy1-binding sites (BS) on yy1a intron (N) and the retinal ChIP assays confirm the loss of Yy1 binding onto its intron, in 2dpi retina (O). (P) The qPCR analysis shows the downregulation of yy1a and yy1b in their own combined knockdown. (Q and R) The schematic showing Yy1-binding sites (BS) on zic2b promoter (Q) and the retinal ChIP assays confirm the physical binding of Yy1 on zic2b promoter, in 2dpi retina (R). (S) Luciferase assay done in zebrafish embryos at 24hpf, injected with zic2b: GFP-luciferase construct shows upregulation in the zic2b promoter activity in yy1a and yy1b knockdown. (T and U) The schematic showing Yy1-binding sites (BS) on oct4 promoter (T) and the retinal ChIP assays confirm the physical binding of Yy1 on oct4 promoter, in 2dpi retina (U). (V) Luciferase assay done in zebrafish embryos at 24hpf, injected with oct4: GFP-luciferase construct shows upregulation in the oct4 promoter activity in yy1a and yy1b knockdown. Error bars represent SD. *p < 0.001 in (H, L, M, P, S, V); *p < 0.05 (F). n = 6 biological replicates (C, E and P).

    Techniques Used: Electroporation, Microscopy, Control, Injection, Western Blot, Knockdown, Binding Assay, Negative Control, Luciferase, Construct, Activity Assay, Over Expression

    (A) An experimental timeline that describes injury, mRNA transfection and electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a overexpression, while no significant increase in the case of yy1b overexpression as compared to control gfp mRNA transfected retina at 4dpi (B), which is quantified (C). (D and E) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a and yy1b combined overexpression, as compared to control gfp mRNA transfected retina at 4dpi (D), which is quantified (E). (F) qPCR analysis shows an upregulation in the levels of ascl1a, lin28a, sox2 and hdac1 in the combined overexpression of yy1a and yy1b. (G) Western blot analysis shows an upregulation in the levels of Ascl1a, Lin28a, Sox2 and Hdac1, while downregulation of Oct4 and Zic2b in the combined overexpression of yy1a and yy1b. (H) qPCR analysis shows a downregulation in the levels of oct4 and zic2b in the combined overexpression of yy1a and yy1b. Scale bars represent 10μm in (B and D); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and D); dpi, days post injury. Error bars represent SD. *p < 0.005 in (E, F); *p < 0.0005 (C, H); n.s is non-significant. n = 6 biological replicates.
    Figure Legend Snippet: (A) An experimental timeline that describes injury, mRNA transfection and electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a overexpression, while no significant increase in the case of yy1b overexpression as compared to control gfp mRNA transfected retina at 4dpi (B), which is quantified (C). (D and E) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a and yy1b combined overexpression, as compared to control gfp mRNA transfected retina at 4dpi (D), which is quantified (E). (F) qPCR analysis shows an upregulation in the levels of ascl1a, lin28a, sox2 and hdac1 in the combined overexpression of yy1a and yy1b. (G) Western blot analysis shows an upregulation in the levels of Ascl1a, Lin28a, Sox2 and Hdac1, while downregulation of Oct4 and Zic2b in the combined overexpression of yy1a and yy1b. (H) qPCR analysis shows a downregulation in the levels of oct4 and zic2b in the combined overexpression of yy1a and yy1b. Scale bars represent 10μm in (B and D); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and D); dpi, days post injury. Error bars represent SD. *p < 0.005 in (E, F); *p < 0.0005 (C, H); n.s is non-significant. n = 6 biological replicates.

    Techniques Used: Transfection, Electroporation, Microscopy, Over Expression, Control, Western Blot

    (A) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug and BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections at 4dpi, show the effect of combined treatment of PFI3 along with yy1a and yy1b overexpression on the number of proliferating cells, which are marked by BrdU + (B), and quantified in (C). (D) qPCR analysis of few RAGs in the experimental conditions explained in B and C. (E) Western blot analysis shows regulation of Ascl1a, Lin28a, Hdac1 and Oct4 in the experimental conditions explained in B and C. (F) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug until 4dpi and BrdU pulse for 5hrs at 4dpi, followed by harvesting at 20dpi. (G and H) IF microscopy images of retinal cross-sections at 20dpi, shows the decrease in number of surviving BrdU positive cells, which were labelled at 4dpi in the combined treatment of PFI3 along with overexpression of yy1a and yy1b mRNA (G), which is quantified in (H). Scale bars represent 10μm in (B and G); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and G); dpi, days post injury. Error bars represent SD. *p < 0.001 in (C, D, H), n.s is non-significant. n = 6 biological replicates.
    Figure Legend Snippet: (A) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug and BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections at 4dpi, show the effect of combined treatment of PFI3 along with yy1a and yy1b overexpression on the number of proliferating cells, which are marked by BrdU + (B), and quantified in (C). (D) qPCR analysis of few RAGs in the experimental conditions explained in B and C. (E) Western blot analysis shows regulation of Ascl1a, Lin28a, Hdac1 and Oct4 in the experimental conditions explained in B and C. (F) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug until 4dpi and BrdU pulse for 5hrs at 4dpi, followed by harvesting at 20dpi. (G and H) IF microscopy images of retinal cross-sections at 20dpi, shows the decrease in number of surviving BrdU positive cells, which were labelled at 4dpi in the combined treatment of PFI3 along with overexpression of yy1a and yy1b mRNA (G), which is quantified in (H). Scale bars represent 10μm in (B and G); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and G); dpi, days post injury. Error bars represent SD. *p < 0.001 in (C, D, H), n.s is non-significant. n = 6 biological replicates.

    Techniques Used: Electroporation, Microscopy, Over Expression, Western Blot



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    rabbit polyclonal antibody against lin28a  (Cell Signaling Technology Inc)
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    Cell Signaling Technology Inc rabbit polyclonal antibody against lin28a
    (A) An experimental timeline that describes injury, Lissamine-tagged Morpholino (MO) delivery, electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a decrease in the number of BrdU + retinal progenitors with yy1a MO and yy1b MO alone and in combination, compared to control MO injected retina at 4dpi (B), which is quantified (C). (D) Western Blot analyses of various RAGs in retinal extracts prepared from retinae electroporated with MOs against yy1a and yy1b at 2dpi. (E) The qPCR analyses of let-7a mi-RNA in the yy1a and yy1b knockdown condition at 2dpi; *p < 0.05; n=6 biological replicates. (F and G) The schematic showing Yy1-binding sites (BS) on ascl1a promoter (F) and the retinal ChIP assays confirm the physical binding of Yy1 on ascl1a promoter, in 2dpi retina (G). N.S marks the negative control. (H and I) Luciferase assay done in zebrafish embryos at 24hpf, injected with ascl1a: GFP-luciferase construct shows upregulation in the ascl1a promoter activity in yy1a and yy1b overexpression (H), while downregulation in their combined knockdown (I). ( J and K) The schematic showing Yy1-binding sites (BS) on <t>lin28a</t> promoter (J) and the retinal ChIP assays confirm the physical binding of Yy1 on lin28a promoter, in 2dpi retina (K). (L and M) Luciferase assay done in zebrafish embryos at 24hpf, injected with lin28a: GFP-luciferase construct shows upregulation in the lin28a promoter activity in yy1a and yy1b overexpression (L), while opposite in their combined knockdown (M). (N and O) The schematic showing Yy1-binding sites (BS) on yy1a intron (N) and the retinal ChIP assays confirm the loss of Yy1 binding onto its intron, in 2dpi retina (O). (P) The qPCR analysis shows the downregulation of yy1a and yy1b in their own combined knockdown. (Q and R) The schematic showing Yy1-binding sites (BS) on zic2b promoter (Q) and the retinal ChIP assays confirm the physical binding of Yy1 on zic2b promoter, in 2dpi retina (R). (S) Luciferase assay done in zebrafish embryos at 24hpf, injected with zic2b: GFP-luciferase construct shows upregulation in the zic2b promoter activity in yy1a and yy1b knockdown. (T and U) The schematic showing Yy1-binding sites (BS) on oct4 promoter (T) and the retinal ChIP assays confirm the physical binding of Yy1 on oct4 promoter, in 2dpi retina (U). (V) Luciferase assay done in zebrafish embryos at 24hpf, injected with oct4: GFP-luciferase construct shows upregulation in the oct4 promoter activity in yy1a and yy1b knockdown. Error bars represent SD. *p < 0.001 in (H, L, M, P, S, V); *p < 0.05 (F). n = 6 biological replicates (C, E and P).
    Rabbit Polyclonal Antibody Against Lin28a, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    rabbit polyclonal antibody against human lin28a  (Proteintech)
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    (A) An experimental timeline that describes injury, Lissamine-tagged Morpholino (MO) delivery, electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a decrease in the number of BrdU + retinal progenitors with yy1a MO and yy1b MO alone and in combination, compared to control MO injected retina at 4dpi (B), which is quantified (C). (D) Western Blot analyses of various RAGs in retinal extracts prepared from retinae electroporated with MOs against yy1a and yy1b at 2dpi. (E) The qPCR analyses of let-7a mi-RNA in the yy1a and yy1b knockdown condition at 2dpi; *p < 0.05; n=6 biological replicates. (F and G) The schematic showing Yy1-binding sites (BS) on ascl1a promoter (F) and the retinal ChIP assays confirm the physical binding of Yy1 on ascl1a promoter, in 2dpi retina (G). N.S marks the negative control. (H and I) Luciferase assay done in zebrafish embryos at 24hpf, injected with ascl1a: GFP-luciferase construct shows upregulation in the ascl1a promoter activity in yy1a and yy1b overexpression (H), while downregulation in their combined knockdown (I). ( J and K) The schematic showing Yy1-binding sites (BS) on <t>lin28a</t> promoter (J) and the retinal ChIP assays confirm the physical binding of Yy1 on lin28a promoter, in 2dpi retina (K). (L and M) Luciferase assay done in zebrafish embryos at 24hpf, injected with lin28a: GFP-luciferase construct shows upregulation in the lin28a promoter activity in yy1a and yy1b overexpression (L), while opposite in their combined knockdown (M). (N and O) The schematic showing Yy1-binding sites (BS) on yy1a intron (N) and the retinal ChIP assays confirm the loss of Yy1 binding onto its intron, in 2dpi retina (O). (P) The qPCR analysis shows the downregulation of yy1a and yy1b in their own combined knockdown. (Q and R) The schematic showing Yy1-binding sites (BS) on zic2b promoter (Q) and the retinal ChIP assays confirm the physical binding of Yy1 on zic2b promoter, in 2dpi retina (R). (S) Luciferase assay done in zebrafish embryos at 24hpf, injected with zic2b: GFP-luciferase construct shows upregulation in the zic2b promoter activity in yy1a and yy1b knockdown. (T and U) The schematic showing Yy1-binding sites (BS) on oct4 promoter (T) and the retinal ChIP assays confirm the physical binding of Yy1 on oct4 promoter, in 2dpi retina (U). (V) Luciferase assay done in zebrafish embryos at 24hpf, injected with oct4: GFP-luciferase construct shows upregulation in the oct4 promoter activity in yy1a and yy1b knockdown. Error bars represent SD. *p < 0.001 in (H, L, M, P, S, V); *p < 0.05 (F). n = 6 biological replicates (C, E and P).
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    Image Search Results


    (A) An experimental timeline that describes injury, Lissamine-tagged Morpholino (MO) delivery, electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a decrease in the number of BrdU + retinal progenitors with yy1a MO and yy1b MO alone and in combination, compared to control MO injected retina at 4dpi (B), which is quantified (C). (D) Western Blot analyses of various RAGs in retinal extracts prepared from retinae electroporated with MOs against yy1a and yy1b at 2dpi. (E) The qPCR analyses of let-7a mi-RNA in the yy1a and yy1b knockdown condition at 2dpi; *p < 0.05; n=6 biological replicates. (F and G) The schematic showing Yy1-binding sites (BS) on ascl1a promoter (F) and the retinal ChIP assays confirm the physical binding of Yy1 on ascl1a promoter, in 2dpi retina (G). N.S marks the negative control. (H and I) Luciferase assay done in zebrafish embryos at 24hpf, injected with ascl1a: GFP-luciferase construct shows upregulation in the ascl1a promoter activity in yy1a and yy1b overexpression (H), while downregulation in their combined knockdown (I). ( J and K) The schematic showing Yy1-binding sites (BS) on lin28a promoter (J) and the retinal ChIP assays confirm the physical binding of Yy1 on lin28a promoter, in 2dpi retina (K). (L and M) Luciferase assay done in zebrafish embryos at 24hpf, injected with lin28a: GFP-luciferase construct shows upregulation in the lin28a promoter activity in yy1a and yy1b overexpression (L), while opposite in their combined knockdown (M). (N and O) The schematic showing Yy1-binding sites (BS) on yy1a intron (N) and the retinal ChIP assays confirm the loss of Yy1 binding onto its intron, in 2dpi retina (O). (P) The qPCR analysis shows the downregulation of yy1a and yy1b in their own combined knockdown. (Q and R) The schematic showing Yy1-binding sites (BS) on zic2b promoter (Q) and the retinal ChIP assays confirm the physical binding of Yy1 on zic2b promoter, in 2dpi retina (R). (S) Luciferase assay done in zebrafish embryos at 24hpf, injected with zic2b: GFP-luciferase construct shows upregulation in the zic2b promoter activity in yy1a and yy1b knockdown. (T and U) The schematic showing Yy1-binding sites (BS) on oct4 promoter (T) and the retinal ChIP assays confirm the physical binding of Yy1 on oct4 promoter, in 2dpi retina (U). (V) Luciferase assay done in zebrafish embryos at 24hpf, injected with oct4: GFP-luciferase construct shows upregulation in the oct4 promoter activity in yy1a and yy1b knockdown. Error bars represent SD. *p < 0.001 in (H, L, M, P, S, V); *p < 0.05 (F). n = 6 biological replicates (C, E and P).

    Journal: bioRxiv

    Article Title: YY1 protein is essential for the promotion of Muller glia reprogramming and retina regeneration

    doi: 10.1101/2025.01.31.635905

    Figure Lengend Snippet: (A) An experimental timeline that describes injury, Lissamine-tagged Morpholino (MO) delivery, electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a decrease in the number of BrdU + retinal progenitors with yy1a MO and yy1b MO alone and in combination, compared to control MO injected retina at 4dpi (B), which is quantified (C). (D) Western Blot analyses of various RAGs in retinal extracts prepared from retinae electroporated with MOs against yy1a and yy1b at 2dpi. (E) The qPCR analyses of let-7a mi-RNA in the yy1a and yy1b knockdown condition at 2dpi; *p < 0.05; n=6 biological replicates. (F and G) The schematic showing Yy1-binding sites (BS) on ascl1a promoter (F) and the retinal ChIP assays confirm the physical binding of Yy1 on ascl1a promoter, in 2dpi retina (G). N.S marks the negative control. (H and I) Luciferase assay done in zebrafish embryos at 24hpf, injected with ascl1a: GFP-luciferase construct shows upregulation in the ascl1a promoter activity in yy1a and yy1b overexpression (H), while downregulation in their combined knockdown (I). ( J and K) The schematic showing Yy1-binding sites (BS) on lin28a promoter (J) and the retinal ChIP assays confirm the physical binding of Yy1 on lin28a promoter, in 2dpi retina (K). (L and M) Luciferase assay done in zebrafish embryos at 24hpf, injected with lin28a: GFP-luciferase construct shows upregulation in the lin28a promoter activity in yy1a and yy1b overexpression (L), while opposite in their combined knockdown (M). (N and O) The schematic showing Yy1-binding sites (BS) on yy1a intron (N) and the retinal ChIP assays confirm the loss of Yy1 binding onto its intron, in 2dpi retina (O). (P) The qPCR analysis shows the downregulation of yy1a and yy1b in their own combined knockdown. (Q and R) The schematic showing Yy1-binding sites (BS) on zic2b promoter (Q) and the retinal ChIP assays confirm the physical binding of Yy1 on zic2b promoter, in 2dpi retina (R). (S) Luciferase assay done in zebrafish embryos at 24hpf, injected with zic2b: GFP-luciferase construct shows upregulation in the zic2b promoter activity in yy1a and yy1b knockdown. (T and U) The schematic showing Yy1-binding sites (BS) on oct4 promoter (T) and the retinal ChIP assays confirm the physical binding of Yy1 on oct4 promoter, in 2dpi retina (U). (V) Luciferase assay done in zebrafish embryos at 24hpf, injected with oct4: GFP-luciferase construct shows upregulation in the oct4 promoter activity in yy1a and yy1b knockdown. Error bars represent SD. *p < 0.001 in (H, L, M, P, S, V); *p < 0.05 (F). n = 6 biological replicates (C, E and P).

    Article Snippet: Primary antibodies used in the study are Rb Anti-YY1, rabbit polyclonal antibody against human ASCL1/MASH1 (Abcam, cat. no. ab74065), Rabbit polyclonal antibody against LIN28a (Cell Signalling Technologies, cat. no. 3978), mouse monoclonal against Oct3/4 (sc5279; Santa Cruz Biotechnology), Mouse polyclonal antibody against Zic2b (raised in house in mice against full length zebrafish Zic2b protein as antigen), rabbit polyclonal antibody against Sox2 (cat. no. ab59776; Abcam), Rabbit polyclonal antibody against pSmad3 (Abcam, ab52903), Rabbit monoclonal antibody against Tgfbi (Abcam, ab170874).

    Techniques: Electroporation, Microscopy, Control, Injection, Western Blot, Knockdown, Binding Assay, Negative Control, Luciferase, Construct, Activity Assay, Over Expression

    (A) An experimental timeline that describes injury, mRNA transfection and electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a overexpression, while no significant increase in the case of yy1b overexpression as compared to control gfp mRNA transfected retina at 4dpi (B), which is quantified (C). (D and E) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a and yy1b combined overexpression, as compared to control gfp mRNA transfected retina at 4dpi (D), which is quantified (E). (F) qPCR analysis shows an upregulation in the levels of ascl1a, lin28a, sox2 and hdac1 in the combined overexpression of yy1a and yy1b. (G) Western blot analysis shows an upregulation in the levels of Ascl1a, Lin28a, Sox2 and Hdac1, while downregulation of Oct4 and Zic2b in the combined overexpression of yy1a and yy1b. (H) qPCR analysis shows a downregulation in the levels of oct4 and zic2b in the combined overexpression of yy1a and yy1b. Scale bars represent 10μm in (B and D); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and D); dpi, days post injury. Error bars represent SD. *p < 0.005 in (E, F); *p < 0.0005 (C, H); n.s is non-significant. n = 6 biological replicates.

    Journal: bioRxiv

    Article Title: YY1 protein is essential for the promotion of Muller glia reprogramming and retina regeneration

    doi: 10.1101/2025.01.31.635905

    Figure Lengend Snippet: (A) An experimental timeline that describes injury, mRNA transfection and electroporation at 0dpi, BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a overexpression, while no significant increase in the case of yy1b overexpression as compared to control gfp mRNA transfected retina at 4dpi (B), which is quantified (C). (D and E) IF microscopy images of retinal cross-sections show a significant increase in the number of BrdU + and PCNA + retinal progenitors in yy1a and yy1b combined overexpression, as compared to control gfp mRNA transfected retina at 4dpi (D), which is quantified (E). (F) qPCR analysis shows an upregulation in the levels of ascl1a, lin28a, sox2 and hdac1 in the combined overexpression of yy1a and yy1b. (G) Western blot analysis shows an upregulation in the levels of Ascl1a, Lin28a, Sox2 and Hdac1, while downregulation of Oct4 and Zic2b in the combined overexpression of yy1a and yy1b. (H) qPCR analysis shows a downregulation in the levels of oct4 and zic2b in the combined overexpression of yy1a and yy1b. Scale bars represent 10μm in (B and D); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and D); dpi, days post injury. Error bars represent SD. *p < 0.005 in (E, F); *p < 0.0005 (C, H); n.s is non-significant. n = 6 biological replicates.

    Article Snippet: Primary antibodies used in the study are Rb Anti-YY1, rabbit polyclonal antibody against human ASCL1/MASH1 (Abcam, cat. no. ab74065), Rabbit polyclonal antibody against LIN28a (Cell Signalling Technologies, cat. no. 3978), mouse monoclonal against Oct3/4 (sc5279; Santa Cruz Biotechnology), Mouse polyclonal antibody against Zic2b (raised in house in mice against full length zebrafish Zic2b protein as antigen), rabbit polyclonal antibody against Sox2 (cat. no. ab59776; Abcam), Rabbit polyclonal antibody against pSmad3 (Abcam, ab52903), Rabbit monoclonal antibody against Tgfbi (Abcam, ab170874).

    Techniques: Transfection, Electroporation, Microscopy, Over Expression, Control, Western Blot

    (A) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug and BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections at 4dpi, show the effect of combined treatment of PFI3 along with yy1a and yy1b overexpression on the number of proliferating cells, which are marked by BrdU + (B), and quantified in (C). (D) qPCR analysis of few RAGs in the experimental conditions explained in B and C. (E) Western blot analysis shows regulation of Ascl1a, Lin28a, Hdac1 and Oct4 in the experimental conditions explained in B and C. (F) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug until 4dpi and BrdU pulse for 5hrs at 4dpi, followed by harvesting at 20dpi. (G and H) IF microscopy images of retinal cross-sections at 20dpi, shows the decrease in number of surviving BrdU positive cells, which were labelled at 4dpi in the combined treatment of PFI3 along with overexpression of yy1a and yy1b mRNA (G), which is quantified in (H). Scale bars represent 10μm in (B and G); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and G); dpi, days post injury. Error bars represent SD. *p < 0.001 in (C, D, H), n.s is non-significant. n = 6 biological replicates.

    Journal: bioRxiv

    Article Title: YY1 protein is essential for the promotion of Muller glia reprogramming and retina regeneration

    doi: 10.1101/2025.01.31.635905

    Figure Lengend Snippet: (A) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug and BrdU pulse for 5hrs at 4dpi, followed by harvesting. (B and C) IF microscopy images of retinal cross-sections at 4dpi, show the effect of combined treatment of PFI3 along with yy1a and yy1b overexpression on the number of proliferating cells, which are marked by BrdU + (B), and quantified in (C). (D) qPCR analysis of few RAGs in the experimental conditions explained in B and C. (E) Western blot analysis shows regulation of Ascl1a, Lin28a, Hdac1 and Oct4 in the experimental conditions explained in B and C. (F) An experimental timeline that describes injury, mRNA delivery, electroporation at 0dpi, dipping in the PFI3 drug until 4dpi and BrdU pulse for 5hrs at 4dpi, followed by harvesting at 20dpi. (G and H) IF microscopy images of retinal cross-sections at 20dpi, shows the decrease in number of surviving BrdU positive cells, which were labelled at 4dpi in the combined treatment of PFI3 along with overexpression of yy1a and yy1b mRNA (G), which is quantified in (H). Scale bars represent 10μm in (B and G); asterisk marks the injury site and GCL, ganglion cell layer; INL, inner nuclear layer; ONL, outer nuclear layer in (B and G); dpi, days post injury. Error bars represent SD. *p < 0.001 in (C, D, H), n.s is non-significant. n = 6 biological replicates.

    Article Snippet: Primary antibodies used in the study are Rb Anti-YY1, rabbit polyclonal antibody against human ASCL1/MASH1 (Abcam, cat. no. ab74065), Rabbit polyclonal antibody against LIN28a (Cell Signalling Technologies, cat. no. 3978), mouse monoclonal against Oct3/4 (sc5279; Santa Cruz Biotechnology), Mouse polyclonal antibody against Zic2b (raised in house in mice against full length zebrafish Zic2b protein as antigen), rabbit polyclonal antibody against Sox2 (cat. no. ab59776; Abcam), Rabbit polyclonal antibody against pSmad3 (Abcam, ab52903), Rabbit monoclonal antibody against Tgfbi (Abcam, ab170874).

    Techniques: Electroporation, Microscopy, Over Expression, Western Blot