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TaKaRa mouse slc39a5 orf sequence

Mouse Slc39a5 Orf Sequence, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/morange+sequence/10__7554_slash_elife__90419-291-0-11?v=TaKaRa
Average 94 stars, based on 225 article reviews

mouse slc39a5 orf sequence - by Bioz Stars, 2026-07

94/100 stars

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1) Product Images from "Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings"

Article Title: Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings

Journal: eLife

doi: 10.7554/elife.90419

Figure Legend Snippet: Figure 1. Rare putative LOF (pLOF) variants in SLC39A5 are associated with elevated serum zinc and nominal protection against type II diabetes (T2D). (A) Serum zinc in carriers of SLC39A5 pLOF variants in the discovery cohort. Controls (Ref; SLC39A5+/+) and heterozygous carriers of pLOF variant alleles in SLC39A5 (Het; SLC39A5+/-). Subject numbers: Ref and Het, respectively: n=5317 and n=15. (B) Trans-ancestry meta-analysis of the association of SLC39A5 pLOF variants with T2D. (C–I) Serum zinc and insulin profile of age, sex and BMI-matched controls in serum call back study. Subject numbers: Ref and Het, respectively: n=246–253 and n=86–91, **p<0.01, unpaired t-test. Numeric data is summarized in Supplementary file 1.

Techniques Used: Variant Assay

Figure 2. Loss of Slc39a5 results in elevated circulating and hepatic zinc levels in mice. Serum zinc (A) and hepatic zinc (B) in Slc39a5+/+, Slc39a5-/-, and Slc39a5+/-mice at 40 wk of age, n=16–18. **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test.

Figure Legend Snippet: Figure 2. Loss of Slc39a5 results in elevated circulating and hepatic zinc levels in mice. Serum zinc (A) and hepatic zinc (B) in Slc39a5+/+, Slc39a5-/-, and Slc39a5+/-mice at 40 wk of age, n=16–18. **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test.

Techniques Used:

Figure 3. Loss of Slc39a5 improves glycemic traits in leptin-receptor deficient mice and in mice challenged with high-fat high fructose diet (HFFD). Female (A-D, I-L; ♀) and Male (E-H, M-P; ♂) mice. (A–H) Slc39a5-/-;Lepr-/- and corresponding control mice. (A, E) Body weight at 34 wk. (B, F) Fasting blood glucose at 34 wk. (C, G) Fasting insulin at 34 wk. (D, H) Homeostatic model assessment for insulin resistance (HOMA-IR) at 34 wk. Slc39a5+/+ and Slc39a5-/- (n=5–12), Lepr -/- and Slc39a5 -/-; Lepr -/- (n=10–15). *p<0.05, **p<0.01, ***p<0.001, one-way ANOVA with post hoc Tukey’s test. (I–P) Slc39a5-/- and Slc39a5+/+ mice were fed HFFD or NC for 30 wk. (I, M) Body weight at 30 wk. (J, N) Fasting blood glucose at 30 wk. (K, O) Fasting insulin at 30 wk. (L, P) HOMA-IR at 30 wk, n=11–15. *p<0.05, **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 4 and Supplementary file 5.

Figure Legend Snippet: Figure 3. Loss of Slc39a5 improves glycemic traits in leptin-receptor deficient mice and in mice challenged with high-fat high fructose diet (HFFD). Female (A-D, I-L; ♀) and Male (E-H, M-P; ♂) mice. (A–H) Slc39a5-/-;Lepr-/- and corresponding control mice. (A, E) Body weight at 34 wk. (B, F) Fasting blood glucose at 34 wk. (C, G) Fasting insulin at 34 wk. (D, H) Homeostatic model assessment for insulin resistance (HOMA-IR) at 34 wk. Slc39a5+/+ and Slc39a5-/- (n=5–12), Lepr -/- and Slc39a5 -/-; Lepr -/- (n=10–15). *p<0.05, **p<0.01, ***p<0.001, one-way ANOVA with post hoc Tukey’s test. (I–P) Slc39a5-/- and Slc39a5+/+ mice were fed HFFD or NC for 30 wk. (I, M) Body weight at 30 wk. (J, N) Fasting blood glucose at 30 wk. (K, O) Fasting insulin at 30 wk. (L, P) HOMA-IR at 30 wk, n=11–15. *p<0.05, **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 4 and Supplementary file 5.

Techniques Used: Control

Figure 4. Loss of Slc39a5 improves liver function and steatosis in leptin-receptor deficient female mice and in female mice challenged with high-fat high fructose diet (HFFD). Slc39a5-/-;Lepr-/- and corresponding control mice (A–F) were sacrificed after 16 hr fasting at 34 wk of age. (G–L) Slc39a5-/- and Slc39a5+/+ mice were fed HFFD or NC for 30 wk and sacrificed after 16 hr of fasting. (A, G) Representative images of livers stained with H&E. Scale bar, 200 µm. (B, H) Hepatic triglyceride (TG) content in explanted liver samples at an endpoint. (C, I) Serum ALT. (D, J) Serum AST. (E, K) Non-alcoholic fatty liver disease (NAFLD) activity score, (F, L) Hepatic beta-hydroxybutyrate (BHOB). *p<0.05, **p<0.01, ***p<0.001, Slc39a5-/-;Lepr-/- and corresponding control mice: one-way ANOVA with post hoc Tukey’s test, HFFD or NC: two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 4 and Supplementary file 5.

Figure Legend Snippet: Figure 4. Loss of Slc39a5 improves liver function and steatosis in leptin-receptor deficient female mice and in female mice challenged with high-fat high fructose diet (HFFD). Slc39a5-/-;Lepr-/- and corresponding control mice (A–F) were sacrificed after 16 hr fasting at 34 wk of age. (G–L) Slc39a5-/- and Slc39a5+/+ mice were fed HFFD or NC for 30 wk and sacrificed after 16 hr of fasting. (A, G) Representative images of livers stained with H&E. Scale bar, 200 µm. (B, H) Hepatic triglyceride (TG) content in explanted liver samples at an endpoint. (C, I) Serum ALT. (D, J) Serum AST. (E, K) Non-alcoholic fatty liver disease (NAFLD) activity score, (F, L) Hepatic beta-hydroxybutyrate (BHOB). *p<0.05, **p<0.01, ***p<0.001, Slc39a5-/-;Lepr-/- and corresponding control mice: one-way ANOVA with post hoc Tukey’s test, HFFD or NC: two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 4 and Supplementary file 5.

Techniques Used: Control, Staining, Activity Assay

Figure 5. Loss of Slc39a5 results in elevated hepatic zinc and activation of hepatic AMPK signaling in leptin-receptor deficient female mice and female mice challenged with high-fat high fructose diet (HFFD). Analyses were done on explanted liver samples collected after 16 hr of fasting at an endpoint in Lepr-/- (A–C) and HFFD mice (D–F). (A, D) Immunoblot analysis of hepatic AMPK and AKT activation. AMPK and AKT signaling is activated in Lepr-/-; Slc39a5-/- mice and HFFD Slc39a5-/- mice (compared to their Scl39a5+/+ counterparts). (B, E) Hepatic zinc is elevated in Lepr-/-; Slc39a5-/- mice and HFFD Slc39a5-/- mice (n=10–21). (C, F) Elevated hepatic zinc results in increased Mt1 (zinc responsive gene) expression in both models. (G) Immunoblot analysis of primary human hepatocytes treated with zinc chloride (ZnCl2), and magnesium chloride (MgCl2), okadaic acid (OA), metformin (Met) for 4 hr. Zinc- activated AMPK and AKT signaling in primary human hepatocytes. (H) Densitometric analysis of immunoblots (compared to control). *p<0.05, **p<0.01, ***p<0.001, ANOVA with post hoc Tukey’s test.

Figure Legend Snippet: Figure 5. Loss of Slc39a5 results in elevated hepatic zinc and activation of hepatic AMPK signaling in leptin-receptor deficient female mice and female mice challenged with high-fat high fructose diet (HFFD). Analyses were done on explanted liver samples collected after 16 hr of fasting at an endpoint in Lepr-/- (A–C) and HFFD mice (D–F). (A, D) Immunoblot analysis of hepatic AMPK and AKT activation. AMPK and AKT signaling is activated in Lepr-/-; Slc39a5-/- mice and HFFD Slc39a5-/- mice (compared to their Scl39a5+/+ counterparts). (B, E) Hepatic zinc is elevated in Lepr-/-; Slc39a5-/- mice and HFFD Slc39a5-/- mice (n=10–21). (C, F) Elevated hepatic zinc results in increased Mt1 (zinc responsive gene) expression in both models. (G) Immunoblot analysis of primary human hepatocytes treated with zinc chloride (ZnCl2), and magnesium chloride (MgCl2), okadaic acid (OA), metformin (Met) for 4 hr. Zinc- activated AMPK and AKT signaling in primary human hepatocytes. (H) Densitometric analysis of immunoblots (compared to control). *p<0.05, **p<0.01, ***p<0.001, ANOVA with post hoc Tukey’s test.

Techniques Used: Activation Assay, Western Blot, Gene Expression, Control

Figure 6. Loss of Slc39a5 improves hepatic inflammation and fibrosis in female mice challenged with diet-induced non-alcoholic steatohepatitis (NASH). Slc39a5-/- and Slc39a5+/+ mice were placed on a NASH-inducing diet or NC for 40 wk and sacrificed after 16 hr of fasting. (A, B) NASH Slc39a5-/- mice display reduced serum ALT and AST levels. (C–E) Histology scores for steatosis, hepatocyte hypertrophy, and inflammation. (F) NAFLD activity score was reduced in NASH Slc39a5-/- mice. (G–I) NASH Slc39a5-/- mice display reduced fibrosis. (G) Representative images of explanted livers sample stained with picrosirius red indicative of collagen deposition. Scale bar, 300 µm. (H, I) Fibrosis and steatosis-activity-fibrosis scores. n=6–7 (NC) and 8–11 (NASH), *p<0.05, **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 6.

Figure Legend Snippet: Figure 6. Loss of Slc39a5 improves hepatic inflammation and fibrosis in female mice challenged with diet-induced non-alcoholic steatohepatitis (NASH). Slc39a5-/- and Slc39a5+/+ mice were placed on a NASH-inducing diet or NC for 40 wk and sacrificed after 16 hr of fasting. (A, B) NASH Slc39a5-/- mice display reduced serum ALT and AST levels. (C–E) Histology scores for steatosis, hepatocyte hypertrophy, and inflammation. (F) NAFLD activity score was reduced in NASH Slc39a5-/- mice. (G–I) NASH Slc39a5-/- mice display reduced fibrosis. (G) Representative images of explanted livers sample stained with picrosirius red indicative of collagen deposition. Scale bar, 300 µm. (H, I) Fibrosis and steatosis-activity-fibrosis scores. n=6–7 (NC) and 8–11 (NASH), *p<0.05, **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 6.

Techniques Used: Activity Assay, Staining

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Journal: eLife

Article Title: Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings

doi: 10.7554/elife.90419

Figure Lengend Snippet: Figure 1. Rare putative LOF (pLOF) variants in SLC39A5 are associated with elevated serum zinc and nominal protection against type II diabetes (T2D). (A) Serum zinc in carriers of SLC39A5 pLOF variants in the discovery cohort. Controls (Ref; SLC39A5+/+) and heterozygous carriers of pLOF variant alleles in SLC39A5 (Het; SLC39A5+/-). Subject numbers: Ref and Het, respectively: n=5317 and n=15. (B) Trans-ancestry meta-analysis of the association of SLC39A5 pLOF variants with T2D. (C–I) Serum zinc and insulin profile of age, sex and BMI-matched controls in serum call back study. Subject numbers: Ref and Het, respectively: n=246–253 and n=86–91, **p<0.01, unpaired t-test. Numeric data is summarized in Supplementary file 1.

Article Snippet: Mouse Slc39a5 ORF sequence was cloned into pIRES2 DsRed- Express2 vector (Clontech, #PT4079- 5).

Techniques: Variant Assay

Journal: eLife

Article Title: Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings

doi: 10.7554/elife.90419

Figure Lengend Snippet: Figure 2. Loss of Slc39a5 results in elevated circulating and hepatic zinc levels in mice. Serum zinc (A) and hepatic zinc (B) in Slc39a5+/+, Slc39a5-/-, and Slc39a5+/-mice at 40 wk of age, n=16–18. **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test.

Article Snippet: Mouse Slc39a5 ORF sequence was cloned into pIRES2 DsRed- Express2 vector (Clontech, #PT4079- 5).

Techniques:

Journal: eLife

Article Title: Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings

doi: 10.7554/elife.90419

Figure Lengend Snippet: Figure 3. Loss of Slc39a5 improves glycemic traits in leptin-receptor deficient mice and in mice challenged with high-fat high fructose diet (HFFD). Female (A-D, I-L; ♀) and Male (E-H, M-P; ♂) mice. (A–H) Slc39a5-/-;Lepr-/- and corresponding control mice. (A, E) Body weight at 34 wk. (B, F) Fasting blood glucose at 34 wk. (C, G) Fasting insulin at 34 wk. (D, H) Homeostatic model assessment for insulin resistance (HOMA-IR) at 34 wk. Slc39a5+/+ and Slc39a5-/- (n=5–12), Lepr -/- and Slc39a5 -/-; Lepr -/- (n=10–15). *p<0.05, **p<0.01, ***p<0.001, one-way ANOVA with post hoc Tukey’s test. (I–P) Slc39a5-/- and Slc39a5+/+ mice were fed HFFD or NC for 30 wk. (I, M) Body weight at 30 wk. (J, N) Fasting blood glucose at 30 wk. (K, O) Fasting insulin at 30 wk. (L, P) HOMA-IR at 30 wk, n=11–15. *p<0.05, **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 4 and Supplementary file 5.

Article Snippet: Mouse Slc39a5 ORF sequence was cloned into pIRES2 DsRed- Express2 vector (Clontech, #PT4079- 5).

Techniques: Control

Journal: eLife

Article Title: Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings

doi: 10.7554/elife.90419

Figure Lengend Snippet: Figure 4. Loss of Slc39a5 improves liver function and steatosis in leptin-receptor deficient female mice and in female mice challenged with high-fat high fructose diet (HFFD). Slc39a5-/-;Lepr-/- and corresponding control mice (A–F) were sacrificed after 16 hr fasting at 34 wk of age. (G–L) Slc39a5-/- and Slc39a5+/+ mice were fed HFFD or NC for 30 wk and sacrificed after 16 hr of fasting. (A, G) Representative images of livers stained with H&E. Scale bar, 200 µm. (B, H) Hepatic triglyceride (TG) content in explanted liver samples at an endpoint. (C, I) Serum ALT. (D, J) Serum AST. (E, K) Non-alcoholic fatty liver disease (NAFLD) activity score, (F, L) Hepatic beta-hydroxybutyrate (BHOB). *p<0.05, **p<0.01, ***p<0.001, Slc39a5-/-;Lepr-/- and corresponding control mice: one-way ANOVA with post hoc Tukey’s test, HFFD or NC: two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 4 and Supplementary file 5.

Article Snippet: Mouse Slc39a5 ORF sequence was cloned into pIRES2 DsRed- Express2 vector (Clontech, #PT4079- 5).

Techniques: Control, Staining, Activity Assay

Journal: eLife

Article Title: Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings

doi: 10.7554/elife.90419

Figure Lengend Snippet: Figure 5. Loss of Slc39a5 results in elevated hepatic zinc and activation of hepatic AMPK signaling in leptin-receptor deficient female mice and female mice challenged with high-fat high fructose diet (HFFD). Analyses were done on explanted liver samples collected after 16 hr of fasting at an endpoint in Lepr-/- (A–C) and HFFD mice (D–F). (A, D) Immunoblot analysis of hepatic AMPK and AKT activation. AMPK and AKT signaling is activated in Lepr-/-; Slc39a5-/- mice and HFFD Slc39a5-/- mice (compared to their Scl39a5+/+ counterparts). (B, E) Hepatic zinc is elevated in Lepr-/-; Slc39a5-/- mice and HFFD Slc39a5-/- mice (n=10–21). (C, F) Elevated hepatic zinc results in increased Mt1 (zinc responsive gene) expression in both models. (G) Immunoblot analysis of primary human hepatocytes treated with zinc chloride (ZnCl2), and magnesium chloride (MgCl2), okadaic acid (OA), metformin (Met) for 4 hr. Zinc- activated AMPK and AKT signaling in primary human hepatocytes. (H) Densitometric analysis of immunoblots (compared to control). *p<0.05, **p<0.01, ***p<0.001, ANOVA with post hoc Tukey’s test.

Article Snippet: Mouse Slc39a5 ORF sequence was cloned into pIRES2 DsRed- Express2 vector (Clontech, #PT4079- 5).

Techniques: Activation Assay, Western Blot, Gene Expression, Control

Journal: eLife

Article Title: Genetic inactivation of zinc transporter SLC39A5 improves liver function and hyperglycemia in obesogenic settings

doi: 10.7554/elife.90419

Figure Lengend Snippet: Figure 6. Loss of Slc39a5 improves hepatic inflammation and fibrosis in female mice challenged with diet-induced non-alcoholic steatohepatitis (NASH). Slc39a5-/- and Slc39a5+/+ mice were placed on a NASH-inducing diet or NC for 40 wk and sacrificed after 16 hr of fasting. (A, B) NASH Slc39a5-/- mice display reduced serum ALT and AST levels. (C–E) Histology scores for steatosis, hepatocyte hypertrophy, and inflammation. (F) NAFLD activity score was reduced in NASH Slc39a5-/- mice. (G–I) NASH Slc39a5-/- mice display reduced fibrosis. (G) Representative images of explanted livers sample stained with picrosirius red indicative of collagen deposition. Scale bar, 300 µm. (H, I) Fibrosis and steatosis-activity-fibrosis scores. n=6–7 (NC) and 8–11 (NASH), *p<0.05, **p<0.01, ***p<0.001, two-way ANOVA with post hoc Tukey’s test. Numeric data is summarized in Supplementary file 6.

Article Snippet: Mouse Slc39a5 ORF sequence was cloned into pIRES2 DsRed- Express2 vector (Clontech, #PT4079- 5).

Techniques: Activity Assay, Staining

A. The location of Pink1 mRNA in MuSCs assessed by FISH assay. Mitochondria were stained by mito-dsRed. White arrows indicate Pink1 mRNAs colocalized with mitochondria. Scale bar, 2μm. B. Quantification of the percentage of Pink1 mRNAs localized in mitochondria in MuSCs based on FISH analysis. C. Schematic of the experimental workflow of identifying Pink1 -associated proteins. D. Silver staining of proteins pulled down by the Pink1 mRNA or Pink1-ATGmut RNA. Negative controls included blank beads and two unrelated RNA sequences. Red arrows indicated proteins specifically bound by Pink1 mRNA or Pink1-ATGmut RNA. E. Ven diagram showing proteins specifically bound by Pink1-ATGmut RNA, categorized based on 3 distinct negative controls. Numbers indicated proteins enriched by Pink1-ATGmut relative to each control. Overlapping areas highlighted proteins that were enriched by Pink1-ATGmut RNA across controls. F. Representative proteins specifically enriched by Pink1-ATGmut RNA shown in panel E, along with their unique peptides and log2 fold enrichment relative to 3 negative controls. Proteins located in mitochondria were colored in purple. G. RNA pulldown followed by Immunoblotting analysis showing the interaction between YME1L1 and Pink1 mRNA. beta-ACTIN serves as the negative control. YME1L1, p: precursor form of YME1L1. YME1L1, m: mature form of YME1L1. Non-specific bands were marked by asterisk. H. Representative images of the mitochondria stained by MitoTracker (red) and DAPI (blue) in scramble (siNC) or Yme1l1 RNAi MuSCs and myotubes, respectively. Scale bars, 4μm and 10μm, respectively. I. Quantification of the mitochondria length in MuSCs and myotubes treated by scramble or Yme1l1 siRNA, respectively. J. Immunoblotting analysis showing OPA1 isoforms in scramble (siNC) and Yme1l1 RNAi (siYme1L1) MuSCs, respectively. Beta-ACTIN served as an internal control. The schematic diagram on the left illustrated the pattern of intermediate products during proteolytic processing of OPA1. S2 indicated the sites cleaved by YME1L1. L-OPA1: long OPA1 isoforms; S-OPA1: short OPA1 isoforms. Yme1l1 knockdown led to insufficient YME1L1 level, which impairs OPA1 processing, leading to accumulation of L-OPA1 and the intermediate proteolytic protein product and decreased S-OPA1 d form. K. Immunoblotting analysis showing OPA1 isoforms in scramble (siNC) and Pink1 RNAi (si Pink1 ) MuSCs, respectively. Beta-ACTIN served as an internal control. L. Immunoblotting analysis showing OPA1 isoforms in WT, Tet2 KO, Tet2 KO with Pink1 overexpressed, and Tet2 KO with Pink1-ATGmut overexpressed MuSCs, respectively. Beta-ACTIN served as an internal control. M. A working model of coding independent function of Pink1 mRNA maintaining mitochondria homeostasis in skeletal muscle cells.

Journal: bioRxiv

Article Title: The non-coding facet of Pink1 mRNA regulates mitochondria homeostasis

doi: 10.64898/2026.01.05.697839

Figure Lengend Snippet: A. The location of Pink1 mRNA in MuSCs assessed by FISH assay. Mitochondria were stained by mito-dsRed. White arrows indicate Pink1 mRNAs colocalized with mitochondria. Scale bar, 2μm. B. Quantification of the percentage of Pink1 mRNAs localized in mitochondria in MuSCs based on FISH analysis. C. Schematic of the experimental workflow of identifying Pink1 -associated proteins. D. Silver staining of proteins pulled down by the Pink1 mRNA or Pink1-ATGmut RNA. Negative controls included blank beads and two unrelated RNA sequences. Red arrows indicated proteins specifically bound by Pink1 mRNA or Pink1-ATGmut RNA. E. Ven diagram showing proteins specifically bound by Pink1-ATGmut RNA, categorized based on 3 distinct negative controls. Numbers indicated proteins enriched by Pink1-ATGmut relative to each control. Overlapping areas highlighted proteins that were enriched by Pink1-ATGmut RNA across controls. F. Representative proteins specifically enriched by Pink1-ATGmut RNA shown in panel E, along with their unique peptides and log2 fold enrichment relative to 3 negative controls. Proteins located in mitochondria were colored in purple. G. RNA pulldown followed by Immunoblotting analysis showing the interaction between YME1L1 and Pink1 mRNA. beta-ACTIN serves as the negative control. YME1L1, p: precursor form of YME1L1. YME1L1, m: mature form of YME1L1. Non-specific bands were marked by asterisk. H. Representative images of the mitochondria stained by MitoTracker (red) and DAPI (blue) in scramble (siNC) or Yme1l1 RNAi MuSCs and myotubes, respectively. Scale bars, 4μm and 10μm, respectively. I. Quantification of the mitochondria length in MuSCs and myotubes treated by scramble or Yme1l1 siRNA, respectively. J. Immunoblotting analysis showing OPA1 isoforms in scramble (siNC) and Yme1l1 RNAi (siYme1L1) MuSCs, respectively. Beta-ACTIN served as an internal control. The schematic diagram on the left illustrated the pattern of intermediate products during proteolytic processing of OPA1. S2 indicated the sites cleaved by YME1L1. L-OPA1: long OPA1 isoforms; S-OPA1: short OPA1 isoforms. Yme1l1 knockdown led to insufficient YME1L1 level, which impairs OPA1 processing, leading to accumulation of L-OPA1 and the intermediate proteolytic protein product and decreased S-OPA1 d form. K. Immunoblotting analysis showing OPA1 isoforms in scramble (siNC) and Pink1 RNAi (si Pink1 ) MuSCs, respectively. Beta-ACTIN served as an internal control. L. Immunoblotting analysis showing OPA1 isoforms in WT, Tet2 KO, Tet2 KO with Pink1 overexpressed, and Tet2 KO with Pink1-ATGmut overexpressed MuSCs, respectively. Beta-ACTIN served as an internal control. M. A working model of coding independent function of Pink1 mRNA maintaining mitochondria homeostasis in skeletal muscle cells.

Article Snippet: Pink1 cDNA fused with a HA tag or a Flag tag at the C terminal, mito-dsRed sequence (Addgene, 44386, 4777bp – 5755bp, US) were cloned into the pLenti-CMV-puro (Addgene, 17448, US) lentiviral expression vector using ClonExpress II One Step Cloning kit (Vazyme, #C112, China), respectively.

Techniques: Staining, Silver Staining, Control, Western Blot, Negative Control, Knockdown

A. The location of Pink1 mRNA in myotubes assessed by FISH assay. Mitochondria were marked by mito-dsRed. White arrows indicate Pink1 mRNAs colocalized with mitochondria. Scale bar, 4μm. B. Quantification of the percentage of Pink1 mRNAs localized in mitochondria in myotubes based on FISH assay. C. RT-qPCR quantification of the mRNA level after scramble (negative control, NC), and Yme1l1 siRNA knockdown (n=3). D. Immunoblotting analysis showing precursor (p) or mature (m) form of YME1L1 in scramble (siNC) and Yme1l1 RNAi (si Yme1l1 ) MuSCs, respectively. GAPDH served as an internal control. E. Immunoblotting analysis showing precursor (p) or mature (m) form of YME1L1 in scramble (siNC) and Pink1 RNAi (si Pink1 ) MuSCs, respectively. Beta-ACTIN served as an internal control. F. Immunoblotting analysis showing precursor (p) or mature (m) form of YME1L1 in WT and Tet2 KO MuSCs, respectively. Beta-ACTIN served as an internal control. G. Dot plot illustrating mRNA abundance rank (the mRNA with the highest abundance ranks 1, the mRNA with the lowest abundance ranks 16,590; mRNAs with same RPM values have the same rank) and protein abundance rank (the protein with the highest abundance ranks 1, the protein with the lowest abundance ranks 8,300, proteins with same abundance have the same rank) for 17,135 protein-coding genes detected either by RNA-seq or spectrometry in WT MuSCs. Black-colored dots indicated genes with discrepant mRNA level and protein level. Red-colored dots indicated genes with high mRNA level but undetectable protein level. Pink1 is highlighted with a green dot.

Journal: bioRxiv

Article Title: The non-coding facet of Pink1 mRNA regulates mitochondria homeostasis

doi: 10.64898/2026.01.05.697839

Figure Lengend Snippet: A. The location of Pink1 mRNA in myotubes assessed by FISH assay. Mitochondria were marked by mito-dsRed. White arrows indicate Pink1 mRNAs colocalized with mitochondria. Scale bar, 4μm. B. Quantification of the percentage of Pink1 mRNAs localized in mitochondria in myotubes based on FISH assay. C. RT-qPCR quantification of the mRNA level after scramble (negative control, NC), and Yme1l1 siRNA knockdown (n=3). D. Immunoblotting analysis showing precursor (p) or mature (m) form of YME1L1 in scramble (siNC) and Yme1l1 RNAi (si Yme1l1 ) MuSCs, respectively. GAPDH served as an internal control. E. Immunoblotting analysis showing precursor (p) or mature (m) form of YME1L1 in scramble (siNC) and Pink1 RNAi (si Pink1 ) MuSCs, respectively. Beta-ACTIN served as an internal control. F. Immunoblotting analysis showing precursor (p) or mature (m) form of YME1L1 in WT and Tet2 KO MuSCs, respectively. Beta-ACTIN served as an internal control. G. Dot plot illustrating mRNA abundance rank (the mRNA with the highest abundance ranks 1, the mRNA with the lowest abundance ranks 16,590; mRNAs with same RPM values have the same rank) and protein abundance rank (the protein with the highest abundance ranks 1, the protein with the lowest abundance ranks 8,300, proteins with same abundance have the same rank) for 17,135 protein-coding genes detected either by RNA-seq or spectrometry in WT MuSCs. Black-colored dots indicated genes with discrepant mRNA level and protein level. Red-colored dots indicated genes with high mRNA level but undetectable protein level. Pink1 is highlighted with a green dot.

Techniques: Quantitative RT-PCR, Negative Control, Knockdown, Western Blot, Control, Quantitative Proteomics, RNA Sequencing

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